immunobloting rubcn (Cell Signaling Technology Inc)

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Cell Signaling Technology Inc immunobloting rubcn

A Variant sparse Partial Least Squares-Discriminate Analysis (sPLS-DA) plot demonstrating group separation among ipsilateral cortical samples from wild-type sham, <t>Rubcn-mutant</t> sham, wild-type injured, and Rubcn-mutant injured mouse groups. B Volcano plots highlighting differentially expressed genes (DEGs) between injured and sham mice in wild-type (left) and Rubcn-mutant (right) samples, with red representing significantly upregulated genes and blue representing significantly downregulated genes. C Volcano plots showing DEGs between Rubcn-mutant and wild-type mice in sham (left) and injured samples (right). A data point corresponding to the Rubcn gene has been removed from the right volcano plot to minimize the y-axis. D Heatmap of gene expression across the samples demonstrating top upregulated (top-half) and downregulated (bottom-half) DEGs between Rubcn-mutant and wild-type TBI mice. E Gene Ontology Biological Process (GO:BP) analysis of significant DEGs (P-value<0.01) between Rubcn-mutant and wild-type TBI samples. Top graph shows GO-terms associated with upregulated genes (in orange) and bottom graph shows GO-terms associated with downregulated genes (in blue) in Rubcn-mutant over wild-type TBI samples with FDR<0.05. F Heat map of selected TBI-induced inflammatory DEG expression across wild-type and Rubcn-mutant sham and TBI samples. G Ingenuity Pathway Analysis (IPA) analysis comparing canonical pathways activated following TBI in wild-type versus Rubcn-mutant mice. Orange (activated) and blue (inhibited) indicate significantly enriched canonical pathways based on activation z-score, while gray dots represent pathways detected in the specified dataset that did not meet significance thresholds. H IPA analysis of lipid-related pathways activated following TBI in wild-type versus Rubcn-mutant mice. Sample size (n) were as follows: WT Sham=5, WT TBI D3=5, Rubcn Mut. Sham=4, Rubcn Mut. TBI D3=6.

Immunobloting Rubcn, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 80 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+rubicon/Rubicon+Rabbit+mAb/bio_rxiv__64898__2026__03__04__709622-165-6-8
Average 95 stars, based on 80 article reviews

immunobloting rubcn - by Bioz Stars, 2026-10

95/100 stars

Images

1) Product Images from "Rubicon modulates neuroimmune responses following traumatic brain injury"

Article Title: Rubicon modulates neuroimmune responses following traumatic brain injury

Journal: bioRxiv

doi: 10.64898/2026.03.04.709622

Figure Legend Snippet: A Variant sparse Partial Least Squares-Discriminate Analysis (sPLS-DA) plot demonstrating group separation among ipsilateral cortical samples from wild-type sham, Rubcn-mutant sham, wild-type injured, and Rubcn-mutant injured mouse groups. B Volcano plots highlighting differentially expressed genes (DEGs) between injured and sham mice in wild-type (left) and Rubcn-mutant (right) samples, with red representing significantly upregulated genes and blue representing significantly downregulated genes. C Volcano plots showing DEGs between Rubcn-mutant and wild-type mice in sham (left) and injured samples (right). A data point corresponding to the Rubcn gene has been removed from the right volcano plot to minimize the y-axis. D Heatmap of gene expression across the samples demonstrating top upregulated (top-half) and downregulated (bottom-half) DEGs between Rubcn-mutant and wild-type TBI mice. E Gene Ontology Biological Process (GO:BP) analysis of significant DEGs (P-value<0.01) between Rubcn-mutant and wild-type TBI samples. Top graph shows GO-terms associated with upregulated genes (in orange) and bottom graph shows GO-terms associated with downregulated genes (in blue) in Rubcn-mutant over wild-type TBI samples with FDR<0.05. F Heat map of selected TBI-induced inflammatory DEG expression across wild-type and Rubcn-mutant sham and TBI samples. G Ingenuity Pathway Analysis (IPA) analysis comparing canonical pathways activated following TBI in wild-type versus Rubcn-mutant mice. Orange (activated) and blue (inhibited) indicate significantly enriched canonical pathways based on activation z-score, while gray dots represent pathways detected in the specified dataset that did not meet significance thresholds. H IPA analysis of lipid-related pathways activated following TBI in wild-type versus Rubcn-mutant mice. Sample size (n) were as follows: WT Sham=5, WT TBI D3=5, Rubcn Mut. Sham=4, Rubcn Mut. TBI D3=6.

Techniques Used: Variant Assay, Mutagenesis, Gene Expression, Expressing, Activation Assay

A Heatmap of RT-qPCR data showing expression of inflammatory genes in ipsilateral cortices of wild-type and Rubcn -mutant mice at 1 dpi (TBI D1) and 3 dpi (TBI D3) in comparison to sham. Color coding is based on z-score. B Fold change in expression levels of indicated genes in wild-type and Rubcn -mutant mice corresponding to A . C Heatmap of RT-qPCR data showing expression of inflammatory genes in ipsilateral cortices of wild-type and Rubcn -mutant mice at 29 dpi (TBI D29). D Fold change in expression levels of indicated genes in wild-type and Rubcn -mutant mice corresponding to C. Bars represent mean ± s.e.m. Sample size ( n ) for A-B were as follows: WT Sham=8, WT TBI D1=5, WT TBI D3=9, Rubcn Mut. Sham=8, Rubcn Mut. D1=7, Rubcn Mut. D3=9. Sample size ( n ) for C-D were as follows: WT Sham=6, WT TBI D29=6, Rubcn Mut. Sham=7, Rubcn Mut. TBI D29=5. Statistical analyses for all data: Two-way ANOVA with post hoc Tukey’s test and significance is represented by * P -value<0.05, ** P -value <0.005, *** P -value <0.0005, **** P -value <0.0001.

Figure Legend Snippet: A Heatmap of RT-qPCR data showing expression of inflammatory genes in ipsilateral cortices of wild-type and Rubcn -mutant mice at 1 dpi (TBI D1) and 3 dpi (TBI D3) in comparison to sham. Color coding is based on z-score. B Fold change in expression levels of indicated genes in wild-type and Rubcn -mutant mice corresponding to A . C Heatmap of RT-qPCR data showing expression of inflammatory genes in ipsilateral cortices of wild-type and Rubcn -mutant mice at 29 dpi (TBI D29). D Fold change in expression levels of indicated genes in wild-type and Rubcn -mutant mice corresponding to C. Bars represent mean ± s.e.m. Sample size ( n ) for A-B were as follows: WT Sham=8, WT TBI D1=5, WT TBI D3=9, Rubcn Mut. Sham=8, Rubcn Mut. D1=7, Rubcn Mut. D3=9. Sample size ( n ) for C-D were as follows: WT Sham=6, WT TBI D29=6, Rubcn Mut. Sham=7, Rubcn Mut. TBI D29=5. Statistical analyses for all data: Two-way ANOVA with post hoc Tukey’s test and significance is represented by * P -value<0.05, ** P -value <0.005, *** P -value <0.0005, **** P -value <0.0001.

Techniques Used: Quantitative RT-PCR, Expressing, Mutagenesis, Comparison

A Number of foot faults before and after injury in wild-type and Rubcn-mutant mice by beam walk test. B Percent of one/two-foot support (left) and three/four-foot support (right) after injury in wild-type and Rubcn-mutant mice by assessed by CatWalk test. C Percentage of ipsilateral cortical damage at 29 dpi between injured wild-type and Rubcn-mutant mice. Data represent mean ± s.e.m. Sample size (n) for A were as follows: WT Sham=11, WT TBI=13, Rubcn Mut. Sham=13, Rubcn Mut. TBI=14. Sample size (n) for B were as follows: n=7-14 mice per group as indicated. Sample size (n) for C were as follows: WT TBI=5, Rubcn Mut. TBI=5. Statistical Analyses: For A-B, mixed-effects analysis with post hoc Tukey’s test; For C, Unpaired Student’s t-test. For all data, significance is represented by * P-value<0.05, ** P-value<0.005.

Figure Legend Snippet: A Number of foot faults before and after injury in wild-type and Rubcn-mutant mice by beam walk test. B Percent of one/two-foot support (left) and three/four-foot support (right) after injury in wild-type and Rubcn-mutant mice by assessed by CatWalk test. C Percentage of ipsilateral cortical damage at 29 dpi between injured wild-type and Rubcn-mutant mice. Data represent mean ± s.e.m. Sample size (n) for A were as follows: WT Sham=11, WT TBI=13, Rubcn Mut. Sham=13, Rubcn Mut. TBI=14. Sample size (n) for B were as follows: n=7-14 mice per group as indicated. Sample size (n) for C were as follows: WT TBI=5, Rubcn Mut. TBI=5. Statistical Analyses: For A-B, mixed-effects analysis with post hoc Tukey’s test; For C, Unpaired Student’s t-test. For all data, significance is represented by * P-value<0.05, ** P-value<0.005.

Techniques Used: Mutagenesis

A Immunoblot comparing damage-associated marker (α-fodrin) and autophagy markers (P62, LC3II) in ipsilateral cortices from wild-type and Rubcn-mutant mice at 3 dpi to sham. B Densitometric quantification of immunoblot in A. C Immunoblot of ipsilateral hippocampus at 1 and 3 dpi in wild-type and Rubcn-mutant mice probed with autophagy markers. D Densitometric quantification of immunoblot in C. E Immunoblot of ipsilateral hippocampus at 1 dpi (TBI D1) and 3 dpi (TBI D3) in wild-type and Rubcn-mutant mice probed with damage-associated markers. F Densitometric quantification of immunoblot in E. G Immunofluorescence (IF) images of GFP-LC3 (green) coronal sections from wild-type and Rubcn-mutant sham and injured mice at 3 dpi stained for IBA1 (in yellow) and P62 (in magenta). Scale bar represents 50 μm. H Quantification of mean fluorescence intensity of P62 (left) and percent of P62/SQSTM1+ cells within LC3+/IBA+ cells (right) from G. All bars represent mean ± s.e.m. Sample size (n) for A-B were as follows: WT Sham=6, WT TBI D3=6, Rubcn Mut. Sham=6, Rubcn Mut. D3=6. Sample size (n) for C-F were as follows: WT Sham=6, WT TBI D1=6, WT TBI D3=6, Rubcn Mut. Sham=5, Rubcn Mut. D1=5, Rubcn Mut. D3=5. Sample size (n) for G-H were as follows: WT Sham=4, WT TBI D3=4, Rubcn Mut. Sham=4, Rubcn Mut. D3=4. Statistical analyses for all data: Two-way ANOVA with post hoc Tukey’s test and significance is represented by * P-value<0.05, ** P-value<0.005, *** P-value<0.0005, **** P-value<0.0001.

Figure Legend Snippet: A Immunoblot comparing damage-associated marker (α-fodrin) and autophagy markers (P62, LC3II) in ipsilateral cortices from wild-type and Rubcn-mutant mice at 3 dpi to sham. B Densitometric quantification of immunoblot in A. C Immunoblot of ipsilateral hippocampus at 1 and 3 dpi in wild-type and Rubcn-mutant mice probed with autophagy markers. D Densitometric quantification of immunoblot in C. E Immunoblot of ipsilateral hippocampus at 1 dpi (TBI D1) and 3 dpi (TBI D3) in wild-type and Rubcn-mutant mice probed with damage-associated markers. F Densitometric quantification of immunoblot in E. G Immunofluorescence (IF) images of GFP-LC3 (green) coronal sections from wild-type and Rubcn-mutant sham and injured mice at 3 dpi stained for IBA1 (in yellow) and P62 (in magenta). Scale bar represents 50 μm. H Quantification of mean fluorescence intensity of P62 (left) and percent of P62/SQSTM1+ cells within LC3+/IBA+ cells (right) from G. All bars represent mean ± s.e.m. Sample size (n) for A-B were as follows: WT Sham=6, WT TBI D3=6, Rubcn Mut. Sham=6, Rubcn Mut. D3=6. Sample size (n) for C-F were as follows: WT Sham=6, WT TBI D1=6, WT TBI D3=6, Rubcn Mut. Sham=5, Rubcn Mut. D1=5, Rubcn Mut. D3=5. Sample size (n) for G-H were as follows: WT Sham=4, WT TBI D3=4, Rubcn Mut. Sham=4, Rubcn Mut. D3=4. Statistical analyses for all data: Two-way ANOVA with post hoc Tukey’s test and significance is represented by * P-value<0.05, ** P-value<0.005, *** P-value<0.0005, **** P-value<0.0001.

Techniques Used: Western Blot, Marker, Mutagenesis, Immunofluorescence, Staining, Fluorescence

A Immunoblot comparing lysates of ipsilateral cortices from wild-type and Rubcn-mutant mice probed with Rubicon A (RbA) antibody. B Western blot showing immunoprecipitation of Rubicon protein from brain lysates of wild-type and Rubcn-mutant mice using Rubicon A (RbA) and Rubicon B (RbA) antibodies. * represents background signal. Blot is probed with RbB antibody. C Rubicon protein (amino-acid) sequence highlighting peptides (p1-p12 in blue) used for peptide mapping of wild-type and mutant RUBCN proteins. Red circle is highlighting Met296 as a potential alternative translation initiation site. D Normalized peptide abundance across the length of RUBCN protein in wild-type and Rubcn-mutant immunoprecipitates using RbA and RbB antibodies. E Comparative enrichment analysis showing fold change of proteins enriched in wild-type over Rubcn-mutant immunoprecipitates using RbA and RbB antibodies. F Immunoprecipitation assay showing lysates of HEK293T cells transfected with indicated plasmids for 24 h and immunoprecipitated with anti-FLAG or anti-HA agarose beads. Left: Immunoblot of immunoprecipitates with the indicated antibodies. Right: Fold change of NRROS signal in EGFP-Rubicon-Flag and Nrros-Myc/His (Nrros+Rubicon1-3) co-transfected cells to EGFP-C1 and Nrros-Myc/His (Nrros ONLY) co-transfected cells. Each FLAG IP sample was normalized to corresponding HA IP sample for quantification. Sample size (n) = 3 biological replicates for EGFP-Rubicon-Flag and Nrros-Myc/His co-transfected cells. G Immunoblot of whole cell lysates used for immunoprecipitation assay in F. H Immunoprecipitation assay showing lysates of HEK293T cells transfected with indicated plasmids for 24 h and immunoprecipitated with anti-MYC or anti-IgG antibodies. Left: Immunoblot of immunoprecipitates with the indicated antibodies. Right: Fold change of RUBCN signal in EGFP-Rubicon-Flag and Nrros-Myc/His (Nrros+Rubicon1-2) co-transfected cells to EGFP-C1 and EGFP-Rubicon-Flag (Rubicon ONLY) co-transfected cells. Each MYC IP sample was normalized to corresponding IgG IP sample. Sample size (n) = 2 biological replicates for EGFP-Rubicon-Flag and Nrros-Myc/His co-transfected cells. I Immunoblot of whole cell lysates used for immunoprecipitation assay in H.

Figure Legend Snippet: A Immunoblot comparing lysates of ipsilateral cortices from wild-type and Rubcn-mutant mice probed with Rubicon A (RbA) antibody. B Western blot showing immunoprecipitation of Rubicon protein from brain lysates of wild-type and Rubcn-mutant mice using Rubicon A (RbA) and Rubicon B (RbA) antibodies. * represents background signal. Blot is probed with RbB antibody. C Rubicon protein (amino-acid) sequence highlighting peptides (p1-p12 in blue) used for peptide mapping of wild-type and mutant RUBCN proteins. Red circle is highlighting Met296 as a potential alternative translation initiation site. D Normalized peptide abundance across the length of RUBCN protein in wild-type and Rubcn-mutant immunoprecipitates using RbA and RbB antibodies. E Comparative enrichment analysis showing fold change of proteins enriched in wild-type over Rubcn-mutant immunoprecipitates using RbA and RbB antibodies. F Immunoprecipitation assay showing lysates of HEK293T cells transfected with indicated plasmids for 24 h and immunoprecipitated with anti-FLAG or anti-HA agarose beads. Left: Immunoblot of immunoprecipitates with the indicated antibodies. Right: Fold change of NRROS signal in EGFP-Rubicon-Flag and Nrros-Myc/His (Nrros+Rubicon1-3) co-transfected cells to EGFP-C1 and Nrros-Myc/His (Nrros ONLY) co-transfected cells. Each FLAG IP sample was normalized to corresponding HA IP sample for quantification. Sample size (n) = 3 biological replicates for EGFP-Rubicon-Flag and Nrros-Myc/His co-transfected cells. G Immunoblot of whole cell lysates used for immunoprecipitation assay in F. H Immunoprecipitation assay showing lysates of HEK293T cells transfected with indicated plasmids for 24 h and immunoprecipitated with anti-MYC or anti-IgG antibodies. Left: Immunoblot of immunoprecipitates with the indicated antibodies. Right: Fold change of RUBCN signal in EGFP-Rubicon-Flag and Nrros-Myc/His (Nrros+Rubicon1-2) co-transfected cells to EGFP-C1 and EGFP-Rubicon-Flag (Rubicon ONLY) co-transfected cells. Each MYC IP sample was normalized to corresponding IgG IP sample. Sample size (n) = 2 biological replicates for EGFP-Rubicon-Flag and Nrros-Myc/His co-transfected cells. I Immunoblot of whole cell lysates used for immunoprecipitation assay in H.

Techniques Used: Western Blot, Mutagenesis, Immunoprecipitation, Sequencing, Transfection

A Immunoblot of ipsilateral cortical tissue lysates from wild-type and Rubcn-mutant mice comparing lipid peroxidation marker at 1 dpi. B Densitometric analysis of immunoblot in A. C Immunoblot of ipsilateral cortical tissue lysates from wild-type and Rubcn-mutant mice comparing lipid peroxidation marker at 3 dpi. D Densitometric analysis of immunoblot in C. Data represent mean ± s.e.m. Sample size (n) were as follows: WT Sham=6, WT TBI D1=6, WT TBI D3=6, Rubcn Mut. Sham=6, Rubcn Mut. D1=6, Rubcn Mut. D3=6.Statistical analyses: Two-way ANOVA with post hoc Tukey’s test and significance is represented by * P-value<0.05, ** P-value<0.005, *** P-value<0.0005, **** P-value<0.0001.

Figure Legend Snippet: A Immunoblot of ipsilateral cortical tissue lysates from wild-type and Rubcn-mutant mice comparing lipid peroxidation marker at 1 dpi. B Densitometric analysis of immunoblot in A. C Immunoblot of ipsilateral cortical tissue lysates from wild-type and Rubcn-mutant mice comparing lipid peroxidation marker at 3 dpi. D Densitometric analysis of immunoblot in C. Data represent mean ± s.e.m. Sample size (n) were as follows: WT Sham=6, WT TBI D1=6, WT TBI D3=6, Rubcn Mut. Sham=6, Rubcn Mut. D1=6, Rubcn Mut. D3=6.Statistical analyses: Two-way ANOVA with post hoc Tukey’s test and significance is represented by * P-value<0.05, ** P-value<0.005, *** P-value<0.0005, **** P-value<0.0001.

Techniques Used: Western Blot, Mutagenesis, Marker

other:

Article Title: Membrane receptors cluster phosphatidylserine to activate LC3-associated phagocytosis
Article Snippet: Lack of TLR2 expression in the negative population was confirmed by western blotting using a different antibody (Cell Signaling Technology, 13744).

Article Title: Elimination of intra-hepatocytic malaria parasites is driven by non-canonical autophagy but not nitric oxide production
Article Snippet: The Western blottings were 801 performed by using iBlot2 PVDF Mini Stacks system (Invitrogen, USA).

Incubation:

Article Title: Membrane receptors cluster phosphatidylserine to activate LC3-associated phagocytosis
Article Snippet: Anti-PS (clone 1H6; EMD-Millipore, 05-719), anti-OSBP8/ORP8 (Abcam, ab99069), anti-CDC50A/TMEM30A (Sigma, AV47410), anti-microtubule-associated protein 1 light chain B (anti-MAPLC3B/LC3; western blots: Cell Signaling Technology, 2775; immunofluorescence: MBL Int, PM036), anti-GABARAP (Cell Signaling Technology, 13733), anti-GABARAPL1 (Cell Signaling Technology, 26632), anti-GABARAPL2 (Abcam, ab126607), anti-sequestosome-1 (anti-SQSTM1/p62; Sigma, P0067), anti-phosphoinositide 3-kinase regulatory subunit 4 (anti-PIK3R4/VPS15; Cell Signaling Technology, 14580), anti-PIK3C3/VPS34 (clone D9A5; Cell Signaling Technology, 4263), anti-Beclin 1 (Cell Signaling Technology, 3738), anti-NADPH oxidase 2 (anti-NOX2/gp91-phox; Santa Cruz Technology, sc-130543), anti-Rubicon (clone D9F7; Cell Signaling Technology, 8465), anti-TLR2 (flow cytometry: clone T2.5, BioLegend, 121810; western blot: clone E1J2W, Cell Signaling Technology, 13744), anti-mCherry (Clonetech, 632543), anti-TREM2 (R&D, MAB17291), anti-LAMP1 (clone 1D4B; Development Studies Hybridoma Bank), anti-ATP6V1A (Proteintech, 17115-1-AP), anti-ATP6V0D1 (Proteintech, 18274-1-AP), anti-ATG5 (Cell Signaling Technology, 12994), anti-ATG7 (Cell Signaling Technology, 8558), anti-ATG16L1 (MBL, M150-3), anti-GST (clone GST-2; Sigma, SAB4200692), anti-FLAG (clone M2; Sigma, F1804), anti-actin (clone C4, horseradish peroxidase (HRP)-conjugated; Santa Cruz, sc-47778), anti-BSA (clone BSA-33; Sigma, B2901) and anti-Biotin (Jackson ImmunoResearch, 200-002-211) were used as indicated in the manufacturers’ datasheets. .. After one wash with 1×PBS, the cells were incubated for 1 h at room temperature with the following primary antibodies diluted in 2%BSA-PBS: anti-FLAG (1:1,000; clone M2; Sigma, F1804), anti-Rubicon (1:500; clone D9F7; Cell Signaling Technology, 8465), anti-LC3B (1:1,000; MBL Int, PM036), anti-VPS34 (1:500; clone D9A5; Cell Signaling Technology, 4263) and anti-LAMP1 (1:200; clone 1D4B; Development Studies Hybridoma Bank). .. The lysis buffer consisted of 50 mM Tris–HCl (pH 7.5), 150 mM NaCl, 5 mM EDTA and 1% (vol/vol) Igepal-CA630 (Sigma, I8896) supplemented with protease inhibitors (cOmplete; Roche, 11836153001) and phosphatase inhibitors (PhosSTOP; Roche, 04906837001).

Article Title: Membrane receptors cluster phosphatidylserine to activate LC3-associated phagocytosis.
Article Snippet: The fixed cells were blocked in 3% (wt/vol) fatty acid-free BSA (3%BSA-PBS; Sigma, 7030) in 1×PBS for 30 min at room temperature. .. After one wash with 1×PBS, the cells were incubated for 1 h at room temperature with the following primary antibodies diluted in 2%BSA-PBS: anti-FLAG (1:1,000; clone M2; Sigma, F1804), anti-Rubicon (1:500; clone D9F7; Cell Signaling Technology, 8465), anti-LC3B (1:1,000; MBL Int, PM036), anti-VPS34 (1:500; clone D9A5; Cell Signaling Technology, 4263) and anti-LAMP1 (1:200; clone 1D4B; Development Studies Hybridoma Bank). ..

Western Blot:

Article Title: Membrane receptors cluster phosphatidylserine to activate LC3-associated phagocytosis.
Article Snippet: The mouse Rubicon cDNA template was a gift from T. Yoshimori (Addgene, 21636). .. Anti-PS (clone 1H6; EMD-Millipore, 05-719), anti-OSBP8/ ORP8 (Abcam, ab99069), anti-CDC50A/TMEM30A (Sigma, AV47410), anti-microtubule-associated protein 1 light chain B (anti-MAPLC3B/ LC3; western blots: Cell Signaling Technology, 2775; immunofluorescence: MBL Int, PM036), anti-GABARAP (Cell Signaling Technology, 13733), anti-GABARAPL1 (Cell Signaling Technology, 26632), anti-GABARAPL2 (Abcam, ab126607), anti-sequestosome-1 (anti-SQSTM1/p62; Sigma, P0067), anti-phosphoinositide 3-kinase regulatory subunit 4 (anti-PIK3R4/VPS15; Cell Signaling Technology, 14580), anti-PIK3C3/VPS34 (clone D9A5; Cell Signaling Technology, 4263), anti-Beclin 1 (Cell Signaling Technology, 3738), anti-NADPH oxidase 2 (anti-NOX2/gp91-phox; Santa Cruz Technology, sc-130543), anti-Rubicon (clone D9F7; Cell Signaling Technology, 8465), anti-TLR2 (flow cytometry: clone T2.5, BioLegend, 121810; western blot: clone E1J2W, Cell Signaling Technology, 13744), anti-mCherry (Clonetech, 632543), anti-TREM2 (R&D, MAB17291), anti-LAMP1 (clone 1D4B; Development Studies Hybridoma Bank), anti-ATP6V1A (Proteintech, 17115-1-AP), anti-ATP6V0D1 (Proteintech, 18274-1-AP), anti-ATG5 (Cell Signaling Technology, 12994), anti-ATG7 (Cell Signaling Technology, 8558), anti-ATG16L1 (MBL, M150-3), anti-GST (clone GST-2; Sigma, SAB4200692), anti-FLAG (clone M2; Sigma, F1804), anti-actin (clone C4, horseradish peroxidase (HRP)-conjugated; Santa Cruz, sc-47778), anti-BSA (clone BSA-33; Sigma, B2901) and anti-Biotin ( Jackson ImmunoResearch, 200-002-211) were used as indicated in the manufacturers’ datasheets. ..

Article Title: Supporting Information for A chemical inhibitor of IST1-CHMP1B interaction impairs endosomal recycling and induces non-canonical LC3 lipidation
Article Snippet: Secondary antibodies were goat anti-rabbit/mouse Alexa Fluor 488 conjugated, goat anti-rabbit/mouse Alexa Fluor 568 conjugated, goat anti-rabbit Alexa Fluor 647 conjugated, goat anti-mouse DyLight 649 conjugated, and all were purchase from Thermo Fisher Scientific. .. Western Blot: anti-IST1 (1:1000, PA5-62981, Invitrogen), anti-Transferrin Receptor (1:1000, 13-6800, Invitrogen), anti-LC3B (1:1000, #2775, Cell Signaling Technology), anti-ATG7 (1:1000, #8558, Cell Signaling Technology), anti-ATG16L1 (1:1000, #8089, Cell Signaling Technology), anti-FIP200 (1:1000, #12436, Cell Signaling Technology), anti-ATG9A (1:1000, #13509, Cell Signaling Technology), anti-Rubicon (1:1000, #8465, Cell Signaling Technology), anti-EGFR (1:100, #4267, Cell Signaling Technology), anti-ALIX (1:1000, #634502, BioLegend), anti-TSG101 (1:1000, #HPA006161, Sigma-Aldrich), anti-actin (1:10000, A2228, Sigma-Aldrich), anti-vimentin (1:1000, #5741, Cell Signaling Technology), anti--tubulin (1:1000, MA5-16308, Invitrogen). .. Secondary antibodies were goat anti-rabbit/ mouse HRP conjugated (31460/ 31430, 1:10000, Thermo Fisher Scientific).

Immunofluorescence:

Article Title: RAB5c controls the assembly of non-canonical autophagy machinery to promote phagosome maturation and microbicidal function of macrophages
Article Snippet: .. Anti-RAB5c [Thermo Fisher Scientific, PA5-39408; IB, 1:2,000; Immunofluorescence (IF), 1:300], anti-GAPDH [Cell Signalling Technology (CST), Clone D16H11, 5174T, IB, 1:10,000], anti-β-actin (CST, Clone 8H10D10, 12262, IB, 1:5,000), anti-Microtubule-associated protein 1 light chain B (anti-MAPLC3B/LC3B; CST, 27759, IB, 1:1,000), anti-LC3 (LC3A/LC3B/LC3C; MBL Int., PM036, IF: 1:250), anti-RAB5a (CST, 2143, IB, 1:1,000), anti-RAB5b (Santa Cruz, Clone F-9, sc373725, IB, 1:1,000), anti-EEA1 (CST, Clone C45B10, 3288, IB, 1:1,000), anti-UNC93B (Abcam, 69497, IB, 1:3,000), anti-GM130 (MBL, Clone 5G8, M1793, IB, 1:500), anti-UVRAG (CST, Clone D2Q1Z, 13115, IB, 1:1,000), anti-Phosphatidylinositol 3-kinase catalytic subunit type 3 (anti-PIK3C3/VPS34; Clone D9A5, CST 4263, IB, 1:1,000), anti-Beclin1 (CST, 3738, IB, 1:1,000), anti-Rubicon (Clone D9F7, CST, 8465, IB, 1:1,000), anti-p-p47 phox (S328) (Abcam, 111855, IF, 1:50), anti-p47 phox (Abcam, Clone erp27205, 308256, IB, 1:1,000; IF, 1:250), anti-p-p40 phox (Thr514) (CST, 4311, IB, 1:1,000), anti-p22 phox (Abcam, 75941 IB, 1:1,000), anti-gp91 phox (BD Bioscience, Clone53, 611414, IB, 1:2,000), anti-ERK1/2 (CST, Clone 1375F, 4695, IF, 1:500), anti-p-ERK1/2(Thr202/204) (CST, Clone 197G2, 9106, IF, 1:100), anti-ATP6V0d1 (Abcam, Clone erp18320-38, IB, 1:1,000), anti-ATP6V1A (Abcam, Clone erp19270, 199326, IB, 1:4,000; IF, 1:250), anti-ATG16L1 (MBL, Clone 1F12, M150-3, IB, 1:1,000), anti-ATG5/12 (CST, Clone D5F5U, 12994, IB, 1:1,000), anti-LAMP1 (CST, Clone E5N9Z, #99437, IF, 1:150), Alexa Fluor 647 polyclonal goat anti-rabbit IgG (Thermo Fisher Scientific, A21244, IF: 1:750), Alexa Fluor 488 polyclonal donkey anti-mouse IgG (Thermo Fisher Scientific, A21202, IF: 1:750), Alexa Fluor 647 polyclonal donkey anti-mouse IgG (Thermo Fisher Scientific, A31571, IF: 1:750), HRP-conjugated anti-rabbit IgG (CST, 7074, IB: 1:5,000), and HRP-conjugated anti-mouse IgG (CST, 7076, IB: 1:5,000) were used as indicated in manufactures′ datasheets. ..

Review

immunobloting rubcn (Cell Signaling Technology Inc)

Structured Review

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1) Product Images from "Rubicon modulates neuroimmune responses following traumatic brain injury"

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