Human PDGF‑C Antibody

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.