Detection of Human Phospho-PDGF R beta (Y751) by Western Blot. Western blot shows lysates of human foreskin fibroblast untreated |
(-) or treated (+) with 100 ng/mL Recombinant Human PDGF‑AA, PDGF‑AB, and PDGF‑BB (Catalog # 221-AA, 222-AB, and 220-BB, respectively) for 10 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human Phospho-PDGF R beta (Y751) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1767), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-PDGF R beta (Y751) at approximately 185 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
|Detection of Human Phospho-PDGF R beta (Y751) by Simple WesternTM. Simple Western lane view shows lysates of CCD‑1070Sk human foreskin fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human PDGF‑AA (Catalog # 221-AA) for 10 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-PDGF R beta (Y751) at approximately 274 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human Phospho-PDGF R beta (Y751) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1767). This experiment was conducted under reducing conditions and using the 66-440 kDa separation system.|
PDGF is a major serum mitogen that can exist as a homo or hetero-dimeric protein consisting of disulfide-linked PDGF-A and PDGF‑B chains. The PDGF-AA, PDGF‑BB and PDGF-AB isoforms have been shown to bind to two distinct cell surface PDGF receptors with different affinities. Where as PDGF R alpha binds all three PDGF isoforms with high affinity, PDGF R beta binds PDGF‑BB only with high-affinity. Both PDGF R alpha and PDGF R beta are members of the class III subfamily of receptor tyrosine kinases (RTK) that also includes the receptors for M-CSF, SCF and Flt3 ligand. All class III RTKs are characterized by the presence of five immunoglobulin‑like domains in their extracellular region and a split kinase domain in their intracellular region. PDGF binding induces receptor homo-and hetero‑dimerization and signal transduction. The expression of the alpha and beta receptors is independently regulated in various cell types. Recombinant soluble PDGF R beta binds PDGF with high affinity and is potent PDGF antagonist.
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