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R&D Systems antimouse pgrn antibody
Antimouse Pgrn Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 87 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/antimouse+pgrn+antibody/10__1016_slash_j__apsb__2026__02__011-80-8-14?v=R%26D+Systems
Average 93 stars, based on 87 article reviews

antimouse pgrn antibody - by Bioz Stars, 2026-08

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$Figure 10. The KO of Grn increases the secretion of TDP-43 and LC3-II from MEFs. A, the base sequences of the genome-edited regions of PGRN in WT and KO mice were elucidated by Sanger sequencing. Arrowheads in the WT alignment show the two bases deleted by CRISPR–Cas9 genome editing. B, representative confocal images of PGRN (green) immunostaining of MEFs derived from PGRN WT and KO mice. Nuclei were stained with DAPI (blue). Arrowheads indicate vesicle-like structures. The image surrounded by a white square in WT MEFs was magniﬁed. Scale bar represents 5 μm. C–H, PGRN WT and KO MEFs were cultured for 72 h. The cell, P1, and P2 fractions were prepared as shown in the Experimental procedures section. C, representative immunoblots (PGRN, TDP-43, TSG101, LC3, and α-tubulin) of the cell, P1, and P2 fractions are shown. The dashed square indicates bands of cleaved TDP-43 used for quantiﬁcation. D, the P1 and P2 fractions treated with or without PNGase F were immunoblotted with PGRN and α-tubulin antibodies. Repre- sentative immunoblots of the cell, P1, and P2 fractions are shown (N = 3). E–H, densitometric data on (E) full-length TDP-43, (F) cleaved TDP-43, (G) TSG101, and (H) LC3-II were calculated from immunoblotting results. Relative signal intensities normalized against PGRN KO MEFs are shown. In bar graphs, data are presented as means ± SD (N = 3). * indicates p < 0.05 by the two-tailed unpaired t test. DAPI, 40,6-diamidino-2-phenylindole; LC3-II, microtubule-associated proteins 1A/1B light chain 3B; MEF, mouse embryonic ﬁbroblast; PGRN, progranulin; TDP-43, transactive response DNA-binding protein 43 kDa; TSG101, tumor susceptibility gene 101 protein.$

Journal: The Journal of biological chemistry

Article Title: Dysregulation of the progranulin-driven autophagy-lysosomal pathway mediates secretion of the nuclear protein TDP-43.

doi: 10.1016/j.jbc.2023.105272

Figure Lengend Snippet: Figure 10. The KO of Grn increases the secretion of TDP-43 and LC3-II from MEFs. A, the base sequences of the genome-edited regions of PGRN in WT and KO mice were elucidated by Sanger sequencing. Arrowheads in the WT alignment show the two bases deleted by CRISPR–Cas9 genome editing. B, representative confocal images of PGRN (green) immunostaining of MEFs derived from PGRN WT and KO mice. Nuclei were stained with DAPI (blue). Arrowheads indicate vesicle-like structures. The image surrounded by a white square in WT MEFs was magniﬁed. Scale bar represents 5 μm. C–H, PGRN WT and KO MEFs were cultured for 72 h. The cell, P1, and P2 fractions were prepared as shown in the Experimental procedures section. C, representative immunoblots (PGRN, TDP-43, TSG101, LC3, and α-tubulin) of the cell, P1, and P2 fractions are shown. The dashed square indicates bands of cleaved TDP-43 used for quantiﬁcation. D, the P1 and P2 fractions treated with or without PNGase F were immunoblotted with PGRN and α-tubulin antibodies. Repre- sentative immunoblots of the cell, P1, and P2 fractions are shown (N = 3). E–H, densitometric data on (E) full-length TDP-43, (F) cleaved TDP-43, (G) TSG101, and (H) LC3-II were calculated from immunoblotting results. Relative signal intensities normalized against PGRN KO MEFs are shown. In bar graphs, data are presented as means ± SD (N = 3). * indicates p < 0.05 by the two-tailed unpaired t test. DAPI, 40,6-diamidino-2-phenylindole; LC3-II, microtubule-associated proteins 1A/1B light chain 3B; MEF, mouse embryonic ﬁbroblast; PGRN, progranulin; TDP-43, transactive response DNA-binding protein 43 kDa; TSG101, tumor susceptibility gene 101 protein.

Article Snippet: * indicates p < 0.05 by the two-tailed un 1A/1B light chain 3B; PGRN, progranulin; TDP-43, transactive response DNA-b Biosciences; a polyclonal sheep antimouse PGRN antibody (catalog no.: AF2557, RRID: AB_2114504) from R&D Systems; a monoclonal mouse anti-WIPI2 antibody (catalog no.: MCA5780GA, RRID: AB_10845951) from Bio-Rad; a biotinconjugated horse antimouse IgG antibody (catalog no.: BA2000, RRID: AB_2313581), biotin-conjugated goat anti-rabbit IgG antibody (catalog no.: BA-1000, RRID: AB_2313606), and biotin-conjugated rabbit antisheep IgG antibody (catalog no.: BA-6000, RRID: AB_2336217) from Vector Laboratories; and Alexa Fluor 568-conjugated donkey antimouse IgG (catalog no.: ab175472, RRID: AB_2636996), and Alexa Fluor 488– conjugated donkey antisheep IgG (catalog no.: ab150177, RRID: AB_2801320) from Abcam.

Techniques: Sequencing, CRISPR, Immunostaining, Derivative Assay, Staining, Cell Culture, Western Blot, Two Tailed Test, Binding Assay

Figure 1. Physical interaction between CD68 and granulin E. A, COS7 cells transfected with CD68-GFP were incubated with conditioned media containing AP-tagged granulin peptides. Positive interaction between CD68 and the granulin E was observed after incubation with AP substrate. The scale bar represents 100 μm. B, Myc-His-tagged CD68 and GFP-tagged granulin A through G were transfected in HEK293T. Lysates were incubated with anti-GFP-conjugated beads. After washes, products were analyzed by Western blot using anti-GFP and anti-Myc antibodies as indicated. C, FLAG-tagged PGRN and GFP-tagged CD68 were transfected in HEK293T cells. Lysates were incubated with anti-GFP-conjugated beads. After washes, products were analyzed by Western blot using anti-GFP and anti-FLAG antibodies as indicated. D, conditioned media containing FLAG-tagged PGRN were incubated with beads bound to GFP or GFP-CD68. After washes, products were analyzed by Western blot using anti-GFP and anti-FLAG antibodies as indicated. E, HEK293T cells were transfected with GFP-tagged CD68 and FLAG-tagged PGRN or PGRN-ΔE. Lysates were incubated with anti-FLAG-conjugated beads. After washes, products were analyzed by Western blot using anti-GFP and anti-FLAG antibodies as indicated. AP, alkaline phosphatase; HEK293T, human embryonic kidney 293T cell line; PGRN, progranulin.

Journal: The Journal of biological chemistry

Article Title: Progranulin-derived granulin E and lysosome membrane protein CD68 interact to reciprocally regulate their protein homeostasis.

doi: 10.1016/j.jbc.2022.102348

Figure Lengend Snippet: Figure 1. Physical interaction between CD68 and granulin E. A, COS7 cells transfected with CD68-GFP were incubated with conditioned media containing AP-tagged granulin peptides. Positive interaction between CD68 and the granulin E was observed after incubation with AP substrate. The scale bar represents 100 μm. B, Myc-His-tagged CD68 and GFP-tagged granulin A through G were transfected in HEK293T. Lysates were incubated with anti-GFP-conjugated beads. After washes, products were analyzed by Western blot using anti-GFP and anti-Myc antibodies as indicated. C, FLAG-tagged PGRN and GFP-tagged CD68 were transfected in HEK293T cells. Lysates were incubated with anti-GFP-conjugated beads. After washes, products were analyzed by Western blot using anti-GFP and anti-FLAG antibodies as indicated. D, conditioned media containing FLAG-tagged PGRN were incubated with beads bound to GFP or GFP-CD68. After washes, products were analyzed by Western blot using anti-GFP and anti-FLAG antibodies as indicated. E, HEK293T cells were transfected with GFP-tagged CD68 and FLAG-tagged PGRN or PGRN-ΔE. Lysates were incubated with anti-FLAG-conjugated beads. After washes, products were analyzed by Western blot using anti-GFP and anti-FLAG antibodies as indicated. AP, alkaline phosphatase; HEK293T, human embryonic kidney 293T cell line; PGRN, progranulin.

Article Snippet: The following antibodies were used in this study: rat antimouse CD68 (Bio-Rad; catalog no.: MCA1957), rabbit anti-CD68 (Abcam; catalog no.: 125212), sheep antimouse PGRN (R&D Systems; catalog no.: AF2557), mouse antiMyc (catalog no.: 9E10; Hybridoma Bank), mouse antiGAPDH (Proteintech Group; catalog no.: 60004-1-Ig), rat antimouse LAMP1 (BD Biosciences; catalog no.: 553792), goat anti-CathD (R&D Systems; catalog no.: AF1029) and rabbit anti-IBA-1 (Wako; catalog no.: 01919741), rabbit anti-PGAM1 (Proteintech; catalog no.: 16126-1-AP), and GFP-Trap were from ChromoTek.

Techniques: Transfection, Incubation, Western Blot

Figure 2. CD68 binds PGRN and granulin E via the mucin–proline-rich domain. A, schematic of CD68 domain structure: mucin–praline-rich domain (M + P) or the LAMP-like domain (Lamp-D), and the transmembrane (TM) domain. B, Myc-tagged CD68 constructs containing the M + P or Lamp- D domains were expressed in HEK293T cells with GFP-tagged PGRN, and the lysates were coimmunoprecipitated using anti-Myc-conjugated beads. IP products were analyzed by Western blot and probed with Myc and GFP antibodies as indicated. Full length-CD68 (FL-CD68) is used as a control. C, M + P or the Lamp-D domains of CD68 were fused to the transmembrane domain of PDGFR (pDisplay vector; Invitrogen) and transfected into COS- 7 cells. FL-CD68 is used as a control. Cells were incubated with AP-tagged PGRN and granulin E (100 nM). The scale bar represents 100 μm. AP, alkaline phosphatase; HEK293T, human embryonic kidney 293T cell line; IP, immunoprecipitation; LAMP, lysosomal-associated membrane protein; PDGFR, platelet-derived growth factor receptor; PGRN, progranulin.

Journal: The Journal of biological chemistry

Article Title: Progranulin-derived granulin E and lysosome membrane protein CD68 interact to reciprocally regulate their protein homeostasis.

doi: 10.1016/j.jbc.2022.102348

Figure Lengend Snippet: Figure 2. CD68 binds PGRN and granulin E via the mucin–proline-rich domain. A, schematic of CD68 domain structure: mucin–praline-rich domain (M + P) or the LAMP-like domain (Lamp-D), and the transmembrane (TM) domain. B, Myc-tagged CD68 constructs containing the M + P or Lamp- D domains were expressed in HEK293T cells with GFP-tagged PGRN, and the lysates were coimmunoprecipitated using anti-Myc-conjugated beads. IP products were analyzed by Western blot and probed with Myc and GFP antibodies as indicated. Full length-CD68 (FL-CD68) is used as a control. C, M + P or the Lamp-D domains of CD68 were fused to the transmembrane domain of PDGFR (pDisplay vector; Invitrogen) and transfected into COS- 7 cells. FL-CD68 is used as a control. Cells were incubated with AP-tagged PGRN and granulin E (100 nM). The scale bar represents 100 μm. AP, alkaline phosphatase; HEK293T, human embryonic kidney 293T cell line; IP, immunoprecipitation; LAMP, lysosomal-associated membrane protein; PDGFR, platelet-derived growth factor receptor; PGRN, progranulin.

Techniques: Construct, Western Blot, Control, Plasmid Preparation, Transfection, Incubation, Immunoprecipitation, Membrane, Derivative Assay

Figure 3. CD68 is not required for PGRN lysosomal trafﬁcking. A, WT and Cd68−/−bone marrow–derived macrophages (BMDMs) were stained with anti- cathepsin D (CathD), PGRN, and CD68 antibodies. The scale bar represents 10 μm (inset: 5 μm). B, brain sections from adult WT and Cd68−/−mice were stained with anti-PGRN, LAMP1, and IBA1 antibodies. The scale bar represents 100 μm (inset: 25 μm). Representative images from three mice of each genotype were shown. C, serum PGRN levels in WT and Cd68−/−mice were measured by ELISA. Six mice per genotype were analyzed (n = 6). D, Western blot analysis of PGRN and granulin peptides in spleen lysates of WT and Cd68−/−mice. Six mice per genotype were analyzed (n = 6). PGRN and granulin in- tensities were normalized to GAPDH. LAMP1, lysosomal-associated membrane protein 1; ns, not signiﬁcant; PGRN, progranulin.

Journal: The Journal of biological chemistry

Article Title: Progranulin-derived granulin E and lysosome membrane protein CD68 interact to reciprocally regulate their protein homeostasis.

doi: 10.1016/j.jbc.2022.102348

Figure Lengend Snippet: Figure 3. CD68 is not required for PGRN lysosomal trafﬁcking. A, WT and Cd68−/−bone marrow–derived macrophages (BMDMs) were stained with anti- cathepsin D (CathD), PGRN, and CD68 antibodies. The scale bar represents 10 μm (inset: 5 μm). B, brain sections from adult WT and Cd68−/−mice were stained with anti-PGRN, LAMP1, and IBA1 antibodies. The scale bar represents 100 μm (inset: 25 μm). Representative images from three mice of each genotype were shown. C, serum PGRN levels in WT and Cd68−/−mice were measured by ELISA. Six mice per genotype were analyzed (n = 6). D, Western blot analysis of PGRN and granulin peptides in spleen lysates of WT and Cd68−/−mice. Six mice per genotype were analyzed (n = 6). PGRN and granulin in- tensities were normalized to GAPDH. LAMP1, lysosomal-associated membrane protein 1; ns, not signiﬁcant; PGRN, progranulin.

Techniques: Derivative Assay, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Membrane

Figure 5. PGRN deﬁciency leads to decreased molecular weight of CD68. A, Western blot analysis of CD68 in WT and Grn−/−spleen lysates. CD68 levels were quantiﬁed and normalized to GAPDH. n = 4, **p < 0.01, unpaired t-test. B, WT and Grn−/−BMDM cells were treated with lysosomal inhibitors, baﬁlomycin (50 nM), and chloroquine (250 μM) for 8 h at 37 C. Representative blots from two replicates were shown. C, Western blot analysis of CD68 in primary microglia derived from WT and Grn−/−mice. Results were from three sets of microglia cultured independently from mouse pups of corresponding genotypes (n = 3). ns, not signiﬁcant; unpaired t-test. D, spleen lysates from WT, Grn−/−, and Cd68−/−mice were immunoprecipitated using rat antimouse CD68 antibodies. The beads were then incubated with or without PNGase F. Representative blots from three replicates are shown. Immunoglobulin G (IgG) bands are indicated by arrows. BMDM, bone marrow–derived macrophage; PGRN, progranulin.

Journal: The Journal of biological chemistry

Article Title: Progranulin-derived granulin E and lysosome membrane protein CD68 interact to reciprocally regulate their protein homeostasis.

doi: 10.1016/j.jbc.2022.102348

Figure Lengend Snippet: Figure 5. PGRN deﬁciency leads to decreased molecular weight of CD68. A, Western blot analysis of CD68 in WT and Grn−/−spleen lysates. CD68 levels were quantiﬁed and normalized to GAPDH. n = 4, **p < 0.01, unpaired t-test. B, WT and Grn−/−BMDM cells were treated with lysosomal inhibitors, baﬁlomycin (50 nM), and chloroquine (250 μM) for 8 h at 37 C. Representative blots from two replicates were shown. C, Western blot analysis of CD68 in primary microglia derived from WT and Grn−/−mice. Results were from three sets of microglia cultured independently from mouse pups of corresponding genotypes (n = 3). ns, not signiﬁcant; unpaired t-test. D, spleen lysates from WT, Grn−/−, and Cd68−/−mice were immunoprecipitated using rat antimouse CD68 antibodies. The beads were then incubated with or without PNGase F. Representative blots from three replicates are shown. Immunoglobulin G (IgG) bands are indicated by arrows. BMDM, bone marrow–derived macrophage; PGRN, progranulin.

Techniques: Molecular Weight, Western Blot, Derivative Assay, Cell Culture, Immunoprecipitation, Incubation

Figure 6. PGRN does not affect CD68 localization. A, WT and Grn−/−BMDMs were stained with antibodies against CD68, cathepsin D, and PGRN. The scale bar represents 10 μm (inset: 5 μm). B, deletion of PGRN in RAW264.7 cells leads to the decreased molecular weight of CD68 and increased CD68 levels. CD68 levels were quantiﬁed and normalized to GAPDH. n = 3, *p < 0.05, unpaired t-test. C, control and Grn−/−RAW264.7 cells were ﬁxed, permeabilized, and stained with anti-CD68, cathepsin D, and PGRN antibodies. The scale bar represents 10 μm (inset: 2.5 μm). D, live control and Grn−/−RAW264.7 cells were incubated with rat anti-mouse CD68 antibodies on ice followed by washing, ﬁxation, blocking, and staining with secondary antibody and Hoechst. Representative images from three replicates are shown. The scale bar represents 10 μm. E, cell surface and total levels of CD68 were quantiﬁed by ImageJ for experiments in C and D. n = 3. BMDM, bone marrow–derived macrophage; ns, not signiﬁcant; PGRN, progranulin.

Journal: The Journal of biological chemistry

Article Title: Progranulin-derived granulin E and lysosome membrane protein CD68 interact to reciprocally regulate their protein homeostasis.

doi: 10.1016/j.jbc.2022.102348

Figure Lengend Snippet: Figure 6. PGRN does not affect CD68 localization. A, WT and Grn−/−BMDMs were stained with antibodies against CD68, cathepsin D, and PGRN. The scale bar represents 10 μm (inset: 5 μm). B, deletion of PGRN in RAW264.7 cells leads to the decreased molecular weight of CD68 and increased CD68 levels. CD68 levels were quantiﬁed and normalized to GAPDH. n = 3, *p < 0.05, unpaired t-test. C, control and Grn−/−RAW264.7 cells were ﬁxed, permeabilized, and stained with anti-CD68, cathepsin D, and PGRN antibodies. The scale bar represents 10 μm (inset: 2.5 μm). D, live control and Grn−/−RAW264.7 cells were incubated with rat anti-mouse CD68 antibodies on ice followed by washing, ﬁxation, blocking, and staining with secondary antibody and Hoechst. Representative images from three replicates are shown. The scale bar represents 10 μm. E, cell surface and total levels of CD68 were quantiﬁed by ImageJ for experiments in C and D. n = 3. BMDM, bone marrow–derived macrophage; ns, not signiﬁcant; PGRN, progranulin.

Techniques: Staining, Molecular Weight, Control, Incubation, Blocking Assay, Derivative Assay

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