Mouse PDGF R beta Antibody

Detection of Rat and Mouse PDGF R beta by Western Blot. Western blot shows lysates of rat spinal cord tissue, rat brain tissue, and mouse brain tissue. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Mouse PDGF R beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1042) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for PDGF R beta at approximately 190 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

PDGF R beta in Mouse Embryonic Spinal Cord. PDGF R beta was detected in immersion fixed frozen sections of mouse embryo spinal cord using 5 µg/mL Goat Anti-Mouse PDGF R beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1042) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of Mouse PDGF R beta by Immunohistochemistry Expression of sFlt1 in pericytes at the angiogenic front. a Maximum intensity projections of confocal images from P6 retinas of the Hey1-GFP transgenic reporter mouse model stained for GFP (green), PDGFR beta + (white) and IB4 (red). Images of the first row show enrichment of Hey1-GFP+, PDGFR beta + perivascular cells in the angiogenic front in comparison to mural cells covering the remodeling central plexus around veins (middle row) and arteries (bottom row). Note strong expression of Hey1-GFP reporter in arterial ECs (bottom row). Scale bar, 50 µm. b, Quantitation of Pdgfrb expression by qPCR in P6 PDGFR beta + retinal pericytes sorted based on GFP expression in comparison to whole-retina single-cell suspension (input). Note significant enrichment of Pdgfrb in both (GFP+ and GFP−) pericyte fractions and higher expression in the Hey1-GFP+ subset. Error bars, s.e.m. p-values, Kruskal–Wallis and Dunn’s multiple comparison test. NS, not statistically significant. c Quantitation of sFlt1 expression by qPCR in sorted P6 retinal pericytes in comparison to whole-retina single-cell suspension (input). Note significant enrichment of sFlt1 expression in Hey1-GFP+ pericytes in comparison to input and GFP- pericytes. Error bars, s.e.m. p-values, one-way ANOVA and Tukey’s multiple comparison test. NS, not statistically significant Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29146905), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse PDGF R beta by Immunocytochemistry/Immunofluorescence VCAM1 is involved in the regulation of proliferation and morphological changes in oligodendrocytes.(a) Tissue lysates from 7-day-old NG2-Cre-driven VCAM1 conditional knockout (VCAM1fl/fl; Ng2-Cre) or control (Ctrl) mouse spinal cords or 11-day-old whole brains were immunoblotted with an antibody against MBP, CNPase, VCAM1 or actin. Data are representative of three experiments. (b–d) Antibodies against Ki67 (green) and Olig2 (red) were used for co-staining in 2-day-old spinal cord cross sections. Data are representative. The scale bars indicate 200 μm. The number of Olig2+ cells per one square millimetre was counted. Data were evaluated using Student’s t-test (**P=0.000528; n=19 slices of two independent experiments). The percentage of Ki67+ cells among the Olig2+ cells is shown. Data were evaluated using Student’s t-test (**P=0.00807; n=6 slices of two independent experiments). (e) Antibodies against Ki67 and NG2 were used for co-staining in 2-day-old spinal cord. The percentage of Ki67+ cells among the NG2+ cells per one square millimetre is shown. Data were evaluated using Student’s t-test (*P=0.0275; n=6 slices of two independent experiments). (f–h) Knock-down efficiencies of shRNAs were confirmed by immunoblotting. Data are representative of three experiments. Oligodendrocytes were transfected with VCAM1 shRNA or control, co-cultured with neurons for 2 days, and co-stained with antibodies against NF (green) and NG2 (red). Magnified images of the dotted boxes (i, attached; ii, aligned) are shown below. Data are representative. Scale bar, 100 μm. The percentage of NG2+ cells whose process tips or cell bodies were attached to axons or aligned along axons is shown. Data were evaluated using Student’s t-test (**P=0.00109 (attached cells) or P=0.00982 (aligned cells); n=6 areas of three experiments). (i,k) Oligodendrocytes were cultured on dishes coated with or without recombinant alpha 4 beta 1 integrin in a growth medium containing PDGF and bFGF. Cells were co-stained with an anti-MBP antibody (red) and DAPI (blue) (a) or an anti-PDGFR alpha antibody (green) (b). Data are representative. Scale bar, 100 μm. The percentage of MBP+ cells is shown. Data were evaluated using Student’s t-test (**P=1.14E−10; n=6 areas of two independent experiments). (j,l) Oligodendrocytes transfected with VCAM1#1 or control shRNA were cultured on recombinant alpha 4 beta 1 integrin-coated dishes in the differentiation medium and co-stained with an anti-MBP antibody (red) and DAPI (blue). Data are representative. Scale bar, 100 μm. The percentage of MBP+ cells is shown. Data were evaluated using Student’s t-test (**P=1.82E−21; n=6 areas of two independent experiments). (m–o) Antibodies against Ki67 (green) and O4 (red) were used for co-staining in 7-day-old spinal cord. DAPI staining is also shown. Data are representative. Scale bar, 100 μm. The number of O4+ cells was counted. Data were evaluated using Student’s t-test (**P=5.87E−05 (P7) or P=5.87E−06 (P14); n=12 slices of two independent experiments). The percentage of Ki67− or Ki67+ cells among the O4+ cells is shown. Data were evaluated using Student’s t-test (**P=0.00344 (P7, Ki67−), 0.00344 (P7, Ki67+) or 0.000580 (P14, Ki67+) *P=0.0494 (P14, Ki67−); n=12 slices of two independent experiments). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27876794), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse PDGF R beta by Immunohistochemistry Vascular alterations after intraocular VEGF-A injection. a Morphology of IB4-stained P6 wild-type retinal vessels at 4 h after administration of human VEGF-A165 (0.5 µl at a concentration of 5 μg μl−1). Note blunt appearance of the vessel front after VEGF-A injection but not for vehicle (PBS) control. Scale bar, 200 µm. b Quantitation of sprouts and filopodia at the front of the P6 vessel plexus after injection of VEGF-A165 or vehicle control. Error bars, s.e.m. p-values, Student’s t-test. c PDGFR beta + (green) pericytes are unaffected by short-term VEGF-A administration, whereas VEGFR2 immunosignals (white) are increased in IB4+ (red) ECs (arrowheads). Images shown correspond to insets in a. Scale bar, 100 µm. d Quantitation of VEGFR2 immunosignals intensity in the peripheral plexus of P6 retinas after injection of VEGF-A165 or vehicle control. Error bars, s.e.m. p-values, Student’s t-test. e Confocal images showing increased Esm1 immunostaining (white) in IB4+ (red) ECs in the peripheral plexus (arrowheads) after VEGF-A injection in P6 pups. Scale bar, 200 µm. f VEGF-A165 injection-mediated increase of Esm1 immunosignals (normalized to IB4+ EC area) in the peripheral capillary plexus but not at the edge of the angiogenic front in comparison to PBS-injected controls at P6. Error bars, s.e.m. p-values, Student’s t-test. NS, not statistically significant. g Short-term VEGF-A165 administration leads to clustering of Erg1+ (green) and IB4+ (red) ECs, as indicated, in thick sprout-like structures of P6 retinas. Panels in the center and on the right (scale bar, 20 µm) show higher magnification of the insets on the left (scale bar, 100 µm). Dashed lines in panels on the right outline IB4+ vessels. h Quantitation of EC density in the leading front vessel and emerging sprouts of the P6 angiogenic front after injection of VEGF-A165 or vehicle control. Error bars, s.e.m. p-values, Student’s t-test Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29146905), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of PDGF R beta in Mouse Embryo Brain. Formalin-fixed paraffin-embedded tissue sections of mouse embryo were probed for PDGFRb mRNA (ACD RNAScope Probe, catalog #411388; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using goat anti-mouse PDGFRb polyclonal antibody (R&D Systems catalog # AF1042) at 1.7ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-goat IgG VisUCyte HRP Polymer Antibody (Catalog # VC004) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to developing vasculature.