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rabbit anti-dynll1  (Thermo Fisher)


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    Structured Review

    Thermo Fisher rabbit anti-dynll1
    Antibodies and assay kits
    Rabbit Anti Dynll1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rabbit+anti-dynll1/pmc11793186-3-0-3
    Average 90 stars, based on 1 article reviews
    rabbit anti-dynll1 - by Bioz Stars, 2026-09
    90/100 stars

    Images

    1) Product Images from "Dynll1-PI31 Interaction Enhances Proteolysis Through the Proteasome, Representing a Novel Therapeutic Target for INF2-Related FSGS"

    Article Title: Dynll1-PI31 Interaction Enhances Proteolysis Through the Proteasome, Representing a Novel Therapeutic Target for INF2-Related FSGS

    Journal: Kidney360

    doi: 10.34067/KID.0000000659

    Antibodies and assay kits
    Figure Legend Snippet: Antibodies and assay kits

    Techniques Used: Membrane, Staining, Activity Assay

    Mouse siRNA duplex sequences
    Figure Legend Snippet: Mouse siRNA duplex sequences

    Techniques Used: Control

    INF2 R218Q promotes proteasome-mediated degradation of nephrin through Dynll1-PI31 interaction . (A) Increased endogenous Dynll1-PI31 interaction and corresponding reduced Dynll1-INF2 interaction in glomerular lysates of R218Q transgenic mice ( wt/ki and ki/ki versus wt/wt ), as measured by Co-IP and quantified as the fraction of INF2 or PI31 protein in the Dynll1-pulldown relative to the total amount in the lysates. n =3, * P < 0.05 versus wt/wt . (B and C) The degradation of nephrin protein in CHX-treated wt and R218Q KI (R218Q) podocytes with siRNA-mediated knockdown of Dynll1 or PI31, compared with cells transfected with control siRNA (B), or in cells treated with Ciliobrevin D (50 µ M) or bortezomib (100 nM), compared with cells treated with vehicle (0.3% DMSO; C). After normalizing nephrin to β -actin, the percent degradation of nephrin was calculated as ([CHX 0 -CHX 2h ]/CHX 0 )×100%. n =3, * P < 0.05 versus wt+control siRNA, ^ P < 0.05 versus R218Q+control siRNA . # P < 0.05 versus R218Q+Ciliobrevin D . (D) Immunofluorescent staining of nephrin (red) and PI31 (green) in wt or R218Q podocytes with different treatments. The MFI of nephrin staining per podocyte and the percentage of peripheral membrane that was nephrin-positive (perimeter labeled in blue by CellBrite 650 membrane stain, Supplemental Figure 2B ) were quantified for comparison. n =5 cells with well-spread peripheral membrane per assay×three independent assays=15), * P < 0.05 versus wt+control siRNA or wt+DMSO, ^ P < 0.05 versus R218Q+control siRNA or R218Q+DMSO . (E) Increased total and K48-specific polyubiquitinated proteins in cells with bortezomib treatment shown in immunoblots of ubiquitinated proteins in both cell lysates and GST-S5a UIM-pulldowns. Cell lysates incubated with GST (instead of GST- S5a) served as a control. (F) Percentage of proteasome activity in cells with siRNA-mediated PI31 knockdown (normalized to the mean of cells treated with control siRNA) and in cells pretreated with bortezomib (normalized to the mean of cells treated with DMSO control). CHX, cycloheximide; Co-IP, co-immunoprecipitation; DMSO, dimethyl sulfoxide; Dynll1, dynein light chain 1; GST, glutathione-S-transferase; INF2, inverted formin 2; IP, immunoprecipitation; KI, knockin; MFI, mean fluorescent intensity; PI31, proteasomal inhibitor of 31kD; UIM, ubiquitin-interacting motif; wt, wild-type.
    Figure Legend Snippet: INF2 R218Q promotes proteasome-mediated degradation of nephrin through Dynll1-PI31 interaction . (A) Increased endogenous Dynll1-PI31 interaction and corresponding reduced Dynll1-INF2 interaction in glomerular lysates of R218Q transgenic mice ( wt/ki and ki/ki versus wt/wt ), as measured by Co-IP and quantified as the fraction of INF2 or PI31 protein in the Dynll1-pulldown relative to the total amount in the lysates. n =3, * P < 0.05 versus wt/wt . (B and C) The degradation of nephrin protein in CHX-treated wt and R218Q KI (R218Q) podocytes with siRNA-mediated knockdown of Dynll1 or PI31, compared with cells transfected with control siRNA (B), or in cells treated with Ciliobrevin D (50 µ M) or bortezomib (100 nM), compared with cells treated with vehicle (0.3% DMSO; C). After normalizing nephrin to β -actin, the percent degradation of nephrin was calculated as ([CHX 0 -CHX 2h ]/CHX 0 )×100%. n =3, * P < 0.05 versus wt+control siRNA, ^ P < 0.05 versus R218Q+control siRNA . # P < 0.05 versus R218Q+Ciliobrevin D . (D) Immunofluorescent staining of nephrin (red) and PI31 (green) in wt or R218Q podocytes with different treatments. The MFI of nephrin staining per podocyte and the percentage of peripheral membrane that was nephrin-positive (perimeter labeled in blue by CellBrite 650 membrane stain, Supplemental Figure 2B ) were quantified for comparison. n =5 cells with well-spread peripheral membrane per assay×three independent assays=15), * P < 0.05 versus wt+control siRNA or wt+DMSO, ^ P < 0.05 versus R218Q+control siRNA or R218Q+DMSO . (E) Increased total and K48-specific polyubiquitinated proteins in cells with bortezomib treatment shown in immunoblots of ubiquitinated proteins in both cell lysates and GST-S5a UIM-pulldowns. Cell lysates incubated with GST (instead of GST- S5a) served as a control. (F) Percentage of proteasome activity in cells with siRNA-mediated PI31 knockdown (normalized to the mean of cells treated with control siRNA) and in cells pretreated with bortezomib (normalized to the mean of cells treated with DMSO control). CHX, cycloheximide; Co-IP, co-immunoprecipitation; DMSO, dimethyl sulfoxide; Dynll1, dynein light chain 1; GST, glutathione-S-transferase; INF2, inverted formin 2; IP, immunoprecipitation; KI, knockin; MFI, mean fluorescent intensity; PI31, proteasomal inhibitor of 31kD; UIM, ubiquitin-interacting motif; wt, wild-type.

    Techniques Used: Transgenic Assay, Co-Immunoprecipitation Assay, Knockdown, Transfection, Control, Staining, Membrane, Labeling, Comparison, Western Blot, Incubation, Activity Assay, Immunoprecipitation, Knock-In

    PA-activated proteasomes deplete nephrin in R218Q KI mice. (A) Reduced nephrin protein in PAN of R218Q KI mice ( wt/ki and ki/ki versus wt/wt ) shown by coimmunofluorescent staining of nephrin (red) and the podocyte marker, WT1 (green). The autofluorescent background was kept in blue channel to show the outline of the mouse kidney histology. (B) Representative Western blots showing the expression of PI31, INF2, Dynll1 and the SD proteins, nephrin and synaptopodin, in mouse glomerular lysates. (C) β -actin-normalized nephrin protein in glomerular lysates was compared in mice of different genotypes and treatments ( n =6 mice per group). (D) Proteasome activity was measured in the glomerular lysates and compared among mice with different genotypes and treatments ( n =6). The inhibitory effect of 25 µ M Lactacystin added into the lysates demonstrated the successful performance of the assay (proteasome activity [+Lactacystin/−Lactacystin]×100%). (E) Coimmunofluorescent staining of nephrin (green) and Dynll1 (red) in kidney sections of control and PA-treated mice of different genotypes ( wt/wt, wt/ki, and ki/ki ). Scale bar: 20 µ m. By using the Coloc2 plugin of Fiji software, Dynll1-nephrin colocalization was quantified as the Manders overlap coefficient and the Pearson correlation coefficient in five randomly picked glomeruli per mouse kidney section and the mean of measurements per mouse kidney section were calculated for comparison. n =6. * P < 0.05 versus control mice of the same genotype , ^ P < 0.05 versus PA-treated mice+NS treatment control of the same genotype. NS, normal saline; PA, puromycin aminonucleoside; PAN, puromycin aminonucleoside nephropathy; RFU, relative fluorescent units; SD, slit diaphragm; WT1, Wilms tumor 1.
    Figure Legend Snippet: PA-activated proteasomes deplete nephrin in R218Q KI mice. (A) Reduced nephrin protein in PAN of R218Q KI mice ( wt/ki and ki/ki versus wt/wt ) shown by coimmunofluorescent staining of nephrin (red) and the podocyte marker, WT1 (green). The autofluorescent background was kept in blue channel to show the outline of the mouse kidney histology. (B) Representative Western blots showing the expression of PI31, INF2, Dynll1 and the SD proteins, nephrin and synaptopodin, in mouse glomerular lysates. (C) β -actin-normalized nephrin protein in glomerular lysates was compared in mice of different genotypes and treatments ( n =6 mice per group). (D) Proteasome activity was measured in the glomerular lysates and compared among mice with different genotypes and treatments ( n =6). The inhibitory effect of 25 µ M Lactacystin added into the lysates demonstrated the successful performance of the assay (proteasome activity [+Lactacystin/−Lactacystin]×100%). (E) Coimmunofluorescent staining of nephrin (green) and Dynll1 (red) in kidney sections of control and PA-treated mice of different genotypes ( wt/wt, wt/ki, and ki/ki ). Scale bar: 20 µ m. By using the Coloc2 plugin of Fiji software, Dynll1-nephrin colocalization was quantified as the Manders overlap coefficient and the Pearson correlation coefficient in five randomly picked glomeruli per mouse kidney section and the mean of measurements per mouse kidney section were calculated for comparison. n =6. * P < 0.05 versus control mice of the same genotype , ^ P < 0.05 versus PA-treated mice+NS treatment control of the same genotype. NS, normal saline; PA, puromycin aminonucleoside; PAN, puromycin aminonucleoside nephropathy; RFU, relative fluorescent units; SD, slit diaphragm; WT1, Wilms tumor 1.

    Techniques Used: Staining, Marker, Western Blot, Expressing, Activity Assay, Control, Software, Comparison, Saline, Wilms Tumor Assay

    Schematics. In R218Q KI podocytes, Dynll1 is not sequestrated by the R218Q variant of INF2, enabling the capture of Dynll1 by PI31, promoting the coupling of the dynein cargo (nephrin) with the proteasome. When the proteasomal proteases are activated by PA, more nephrin is degraded, decreasing the pool of nephrin available for surface trafficking. This pathogenic process can be broken by PIs. In wt podocytes, Dynll1 is sequestered by INF2 and the proteasome-mediated degradation of nephrin is limited, allowing adequate flow of nephrin for functional targeting to the cell surface and subsequent incorporation into SDs. PIs, proteasome inhibitors.
    Figure Legend Snippet: Schematics. In R218Q KI podocytes, Dynll1 is not sequestrated by the R218Q variant of INF2, enabling the capture of Dynll1 by PI31, promoting the coupling of the dynein cargo (nephrin) with the proteasome. When the proteasomal proteases are activated by PA, more nephrin is degraded, decreasing the pool of nephrin available for surface trafficking. This pathogenic process can be broken by PIs. In wt podocytes, Dynll1 is sequestered by INF2 and the proteasome-mediated degradation of nephrin is limited, allowing adequate flow of nephrin for functional targeting to the cell surface and subsequent incorporation into SDs. PIs, proteasome inhibitors.

    Techniques Used: Variant Assay, Functional Assay

    Related Articles

    Membrane:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Staining:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Activity Assay:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Control:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Transgenic Assay:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Co-Immunoprecipitation Assay:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Knockdown:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Transfection:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Labeling:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Comparison:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Western Blot:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Incubation:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Immunoprecipitation:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Knock-In:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Marker:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Expressing:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Software:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Saline:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Wilms Tumor Assay:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Variant Assay:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Functional Assay:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Histone Deacetylase Assay:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    In Vitro:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Immunofluorescence:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Synthesized:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Stripping Membranes:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Live Cell Imaging:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    In Vivo:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Surface Biotinylation Assay:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    In Situ:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.



    Biomarker Assay:

    Article Title: Dynein-Mediated Trafficking: A New Mechanism of Diabetic Podocytopathy
    Article Snippet: Table 2 Antibody Dilution Company Cat# Mouse antinephrin (G-8) IF: 1:100 Santa Cruz sc-376522 Mouse antinephrin conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-376522 AC Normal mouse IgG conjugated to agarose IP: 2 μg/400 μl lysate Santa Cruz sc-2343 Rabbit antinephrin WB: 1:1000 Invitrogen PA5-91907 Sheep anti-DCTN1 WB: 1:1000 IF: 1:100 R&D Systems AF6657 Mouse anti-Dynll1 IF: 1:100 Santa Cruz sc-136287 Rabbit anti-Dynll1 WB: 1:1000 IF: 1:100 Thermo Fisher PA5-97920 Rabbit anti-INF2 IF: 1:100 Bethyl Lab A303-427A Rabbit anti-HDAC6 WB: 1:1000 Novus NBP1-78981 Donkey anti-mouse (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32766 Donkey anti-rabbit (Alexa Fluor 488) IF: 1:200 Thermo Fisher A32790 Donkey anti-mouse (Alexa Fluor 594) IF: 1:200 Thermo Fisher A32744 Donkey anti-sheep (Alexa Fluor 488) IF: 1:200 Thermo Fisher A-11015 Mouse anti-rabbit IgG-HRP WB: 1:5000 Santa Cruz sc-2357 Rabbit anti-sheep IgG-HRP WB: 1:5000 Thermo Fisher 31480 Biotin mouse anti-ubiquitin WB: 1:1000 Invitrogen 13-6078-82 Mouse anti β-actin-HRP WB: 1:5000 Santa Cruz sc-47778 Mouse anti-WT1 IF: 1:100 Novus NBP24460700 Mouse anti-acetylated α-tubulin IF: 1:200 Santa Cruz sc-23950 Acti-stain 488 phalloidin Cytoskeleton PHDG1-A ViaFluor microtubule stains (488) Biotium 70062 Open in a separate window IF, immunofluorescent; IP, immunoprecipitation; WB, Western blotting; INF2, inverted formin 2; HDAC6, histone deacetylase 6; HRP, horseradish peroxidase.





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    Image Search Results


    Antibodies and assay kits

    Journal: Kidney360

    Article Title: Dynll1-PI31 Interaction Enhances Proteolysis Through the Proteasome, Representing a Novel Therapeutic Target for INF2-Related FSGS

    doi: 10.34067/KID.0000000659

    Figure Lengend Snippet: Antibodies and assay kits

    Article Snippet: Rabbit anti-Dynll1 , Thermo Fisher , PA5-97920.

    Techniques: Membrane, Staining, Activity Assay

    Mouse siRNA duplex sequences

    Journal: Kidney360

    Article Title: Dynll1-PI31 Interaction Enhances Proteolysis Through the Proteasome, Representing a Novel Therapeutic Target for INF2-Related FSGS

    doi: 10.34067/KID.0000000659

    Figure Lengend Snippet: Mouse siRNA duplex sequences

    Article Snippet: Rabbit anti-Dynll1 , Thermo Fisher , PA5-97920.

    Techniques: Control

    INF2 R218Q promotes proteasome-mediated degradation of nephrin through Dynll1-PI31 interaction . (A) Increased endogenous Dynll1-PI31 interaction and corresponding reduced Dynll1-INF2 interaction in glomerular lysates of R218Q transgenic mice ( wt/ki and ki/ki versus wt/wt ), as measured by Co-IP and quantified as the fraction of INF2 or PI31 protein in the Dynll1-pulldown relative to the total amount in the lysates. n =3, * P < 0.05 versus wt/wt . (B and C) The degradation of nephrin protein in CHX-treated wt and R218Q KI (R218Q) podocytes with siRNA-mediated knockdown of Dynll1 or PI31, compared with cells transfected with control siRNA (B), or in cells treated with Ciliobrevin D (50 µ M) or bortezomib (100 nM), compared with cells treated with vehicle (0.3% DMSO; C). After normalizing nephrin to β -actin, the percent degradation of nephrin was calculated as ([CHX 0 -CHX 2h ]/CHX 0 )×100%. n =3, * P < 0.05 versus wt+control siRNA, ^ P < 0.05 versus R218Q+control siRNA . # P < 0.05 versus R218Q+Ciliobrevin D . (D) Immunofluorescent staining of nephrin (red) and PI31 (green) in wt or R218Q podocytes with different treatments. The MFI of nephrin staining per podocyte and the percentage of peripheral membrane that was nephrin-positive (perimeter labeled in blue by CellBrite 650 membrane stain, Supplemental Figure 2B ) were quantified for comparison. n =5 cells with well-spread peripheral membrane per assay×three independent assays=15), * P < 0.05 versus wt+control siRNA or wt+DMSO, ^ P < 0.05 versus R218Q+control siRNA or R218Q+DMSO . (E) Increased total and K48-specific polyubiquitinated proteins in cells with bortezomib treatment shown in immunoblots of ubiquitinated proteins in both cell lysates and GST-S5a UIM-pulldowns. Cell lysates incubated with GST (instead of GST- S5a) served as a control. (F) Percentage of proteasome activity in cells with siRNA-mediated PI31 knockdown (normalized to the mean of cells treated with control siRNA) and in cells pretreated with bortezomib (normalized to the mean of cells treated with DMSO control). CHX, cycloheximide; Co-IP, co-immunoprecipitation; DMSO, dimethyl sulfoxide; Dynll1, dynein light chain 1; GST, glutathione-S-transferase; INF2, inverted formin 2; IP, immunoprecipitation; KI, knockin; MFI, mean fluorescent intensity; PI31, proteasomal inhibitor of 31kD; UIM, ubiquitin-interacting motif; wt, wild-type.

    Journal: Kidney360

    Article Title: Dynll1-PI31 Interaction Enhances Proteolysis Through the Proteasome, Representing a Novel Therapeutic Target for INF2-Related FSGS

    doi: 10.34067/KID.0000000659

    Figure Lengend Snippet: INF2 R218Q promotes proteasome-mediated degradation of nephrin through Dynll1-PI31 interaction . (A) Increased endogenous Dynll1-PI31 interaction and corresponding reduced Dynll1-INF2 interaction in glomerular lysates of R218Q transgenic mice ( wt/ki and ki/ki versus wt/wt ), as measured by Co-IP and quantified as the fraction of INF2 or PI31 protein in the Dynll1-pulldown relative to the total amount in the lysates. n =3, * P < 0.05 versus wt/wt . (B and C) The degradation of nephrin protein in CHX-treated wt and R218Q KI (R218Q) podocytes with siRNA-mediated knockdown of Dynll1 or PI31, compared with cells transfected with control siRNA (B), or in cells treated with Ciliobrevin D (50 µ M) or bortezomib (100 nM), compared with cells treated with vehicle (0.3% DMSO; C). After normalizing nephrin to β -actin, the percent degradation of nephrin was calculated as ([CHX 0 -CHX 2h ]/CHX 0 )×100%. n =3, * P < 0.05 versus wt+control siRNA, ^ P < 0.05 versus R218Q+control siRNA . # P < 0.05 versus R218Q+Ciliobrevin D . (D) Immunofluorescent staining of nephrin (red) and PI31 (green) in wt or R218Q podocytes with different treatments. The MFI of nephrin staining per podocyte and the percentage of peripheral membrane that was nephrin-positive (perimeter labeled in blue by CellBrite 650 membrane stain, Supplemental Figure 2B ) were quantified for comparison. n =5 cells with well-spread peripheral membrane per assay×three independent assays=15), * P < 0.05 versus wt+control siRNA or wt+DMSO, ^ P < 0.05 versus R218Q+control siRNA or R218Q+DMSO . (E) Increased total and K48-specific polyubiquitinated proteins in cells with bortezomib treatment shown in immunoblots of ubiquitinated proteins in both cell lysates and GST-S5a UIM-pulldowns. Cell lysates incubated with GST (instead of GST- S5a) served as a control. (F) Percentage of proteasome activity in cells with siRNA-mediated PI31 knockdown (normalized to the mean of cells treated with control siRNA) and in cells pretreated with bortezomib (normalized to the mean of cells treated with DMSO control). CHX, cycloheximide; Co-IP, co-immunoprecipitation; DMSO, dimethyl sulfoxide; Dynll1, dynein light chain 1; GST, glutathione-S-transferase; INF2, inverted formin 2; IP, immunoprecipitation; KI, knockin; MFI, mean fluorescent intensity; PI31, proteasomal inhibitor of 31kD; UIM, ubiquitin-interacting motif; wt, wild-type.

    Article Snippet: Rabbit anti-Dynll1 , Thermo Fisher , PA5-97920.

    Techniques: Transgenic Assay, Co-Immunoprecipitation Assay, Knockdown, Transfection, Control, Staining, Membrane, Labeling, Comparison, Western Blot, Incubation, Activity Assay, Immunoprecipitation, Knock-In

    PA-activated proteasomes deplete nephrin in R218Q KI mice. (A) Reduced nephrin protein in PAN of R218Q KI mice ( wt/ki and ki/ki versus wt/wt ) shown by coimmunofluorescent staining of nephrin (red) and the podocyte marker, WT1 (green). The autofluorescent background was kept in blue channel to show the outline of the mouse kidney histology. (B) Representative Western blots showing the expression of PI31, INF2, Dynll1 and the SD proteins, nephrin and synaptopodin, in mouse glomerular lysates. (C) β -actin-normalized nephrin protein in glomerular lysates was compared in mice of different genotypes and treatments ( n =6 mice per group). (D) Proteasome activity was measured in the glomerular lysates and compared among mice with different genotypes and treatments ( n =6). The inhibitory effect of 25 µ M Lactacystin added into the lysates demonstrated the successful performance of the assay (proteasome activity [+Lactacystin/−Lactacystin]×100%). (E) Coimmunofluorescent staining of nephrin (green) and Dynll1 (red) in kidney sections of control and PA-treated mice of different genotypes ( wt/wt, wt/ki, and ki/ki ). Scale bar: 20 µ m. By using the Coloc2 plugin of Fiji software, Dynll1-nephrin colocalization was quantified as the Manders overlap coefficient and the Pearson correlation coefficient in five randomly picked glomeruli per mouse kidney section and the mean of measurements per mouse kidney section were calculated for comparison. n =6. * P < 0.05 versus control mice of the same genotype , ^ P < 0.05 versus PA-treated mice+NS treatment control of the same genotype. NS, normal saline; PA, puromycin aminonucleoside; PAN, puromycin aminonucleoside nephropathy; RFU, relative fluorescent units; SD, slit diaphragm; WT1, Wilms tumor 1.

    Journal: Kidney360

    Article Title: Dynll1-PI31 Interaction Enhances Proteolysis Through the Proteasome, Representing a Novel Therapeutic Target for INF2-Related FSGS

    doi: 10.34067/KID.0000000659

    Figure Lengend Snippet: PA-activated proteasomes deplete nephrin in R218Q KI mice. (A) Reduced nephrin protein in PAN of R218Q KI mice ( wt/ki and ki/ki versus wt/wt ) shown by coimmunofluorescent staining of nephrin (red) and the podocyte marker, WT1 (green). The autofluorescent background was kept in blue channel to show the outline of the mouse kidney histology. (B) Representative Western blots showing the expression of PI31, INF2, Dynll1 and the SD proteins, nephrin and synaptopodin, in mouse glomerular lysates. (C) β -actin-normalized nephrin protein in glomerular lysates was compared in mice of different genotypes and treatments ( n =6 mice per group). (D) Proteasome activity was measured in the glomerular lysates and compared among mice with different genotypes and treatments ( n =6). The inhibitory effect of 25 µ M Lactacystin added into the lysates demonstrated the successful performance of the assay (proteasome activity [+Lactacystin/−Lactacystin]×100%). (E) Coimmunofluorescent staining of nephrin (green) and Dynll1 (red) in kidney sections of control and PA-treated mice of different genotypes ( wt/wt, wt/ki, and ki/ki ). Scale bar: 20 µ m. By using the Coloc2 plugin of Fiji software, Dynll1-nephrin colocalization was quantified as the Manders overlap coefficient and the Pearson correlation coefficient in five randomly picked glomeruli per mouse kidney section and the mean of measurements per mouse kidney section were calculated for comparison. n =6. * P < 0.05 versus control mice of the same genotype , ^ P < 0.05 versus PA-treated mice+NS treatment control of the same genotype. NS, normal saline; PA, puromycin aminonucleoside; PAN, puromycin aminonucleoside nephropathy; RFU, relative fluorescent units; SD, slit diaphragm; WT1, Wilms tumor 1.

    Article Snippet: Rabbit anti-Dynll1 , Thermo Fisher , PA5-97920.

    Techniques: Staining, Marker, Western Blot, Expressing, Activity Assay, Control, Software, Comparison, Saline, Wilms Tumor Assay

    Schematics. In R218Q KI podocytes, Dynll1 is not sequestrated by the R218Q variant of INF2, enabling the capture of Dynll1 by PI31, promoting the coupling of the dynein cargo (nephrin) with the proteasome. When the proteasomal proteases are activated by PA, more nephrin is degraded, decreasing the pool of nephrin available for surface trafficking. This pathogenic process can be broken by PIs. In wt podocytes, Dynll1 is sequestered by INF2 and the proteasome-mediated degradation of nephrin is limited, allowing adequate flow of nephrin for functional targeting to the cell surface and subsequent incorporation into SDs. PIs, proteasome inhibitors.

    Journal: Kidney360

    Article Title: Dynll1-PI31 Interaction Enhances Proteolysis Through the Proteasome, Representing a Novel Therapeutic Target for INF2-Related FSGS

    doi: 10.34067/KID.0000000659

    Figure Lengend Snippet: Schematics. In R218Q KI podocytes, Dynll1 is not sequestrated by the R218Q variant of INF2, enabling the capture of Dynll1 by PI31, promoting the coupling of the dynein cargo (nephrin) with the proteasome. When the proteasomal proteases are activated by PA, more nephrin is degraded, decreasing the pool of nephrin available for surface trafficking. This pathogenic process can be broken by PIs. In wt podocytes, Dynll1 is sequestered by INF2 and the proteasome-mediated degradation of nephrin is limited, allowing adequate flow of nephrin for functional targeting to the cell surface and subsequent incorporation into SDs. PIs, proteasome inhibitors.

    Article Snippet: Rabbit anti-Dynll1 , Thermo Fisher , PA5-97920.

    Techniques: Variant Assay, Functional Assay