Human PDGF-C Antibody

Detection of Human PDGF‑C by Western Blot. Western blot shows lysates of human platelets. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human PDGF-C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1560) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). Specific bands were detected for PDGF-C p80 at approximately 80 kDa and PDGF-C p55 at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

PDGF‑C in Human Pancreatic Cancer Tissue. PDGF‑C was detected in immersion fixed paraffin-embedded sections of human pancreatic cancer tissue using 1.7 µg/mL Goat Anti-Human PDGF‑C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1560) 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 Paraffin-embedded Tissue Sections.

Cell Proliferation Induced by PDGF‑CC and Neutralization by Human PDGF‑C Antibody. Recombinant Human PDGF-CC (Catalog # 1687-CC) stimulates proliferation in the NR6R-3T3 mouse fibroblast cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human PDGF-CC (0.8 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human PDGF-C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1560). The ND₅₀ is typically 6-24 µg/mL.

Detection of Human PDGF-C by Western Blot PDGF-C binds to NRP-1 in vitro and stimulates signal transduction in M14-N cells(A) PDGF-C binding to NRP-1 was assayed in Maxisorp Nunc immunoplates coated with the growth factor and incubated with a solution containing 1 μg/ml NRP-1/hFc chimera. Bound NRP-1/hFc chimeric polypeptide was quantified using an alkaline phosphatase-conjugated anti-human Fc antibody. PDGFR alpha /hFc and VEGFR-2/hFc chimeras were used as positive and negative controls, respectively. Each value represents the mean of three independent determinations (± SD). ANOVA followed by Bonferroni’s post-hoc test: p<0.05 (*). (B) The effect of pre-incubation of selected PDGF-C-coated wells with graded concentrations of goat anti-PDGF-C or IgG control antibodies on PDGF-C binding to NRP-1 was evaluated using the binding assay described in panel A. Student’s t-test: p<0.05 (*), p<0.01 (**). (C) M14-N cells were untreated or treated with 50 ng/ml PDGF-C for the indicated times and p130Cas phosphorylation levels were evaluated by Western blot. Total p130Cas was detected to determine the relative amount of phosphorylated protein and beta -tubulin expression was tested as loading control. Protein extracts from M14-C cells, at time 5 min, were included in the analysis as negative control. A representative experiment out of three is shown. (D) Densitometric quantification of the levels of phospho-p130Cas relative to the total amount of p130Cas, after normalization by beta -tubulin content in the samples. Histogram represents the mean values (± SD) of three independent determinations. Student’s t-test analysis: p<0.01 (**). NS, non-stimulated cells. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28977999), licensed under a CC-BY license. Not internally tested by R&D Systems.