Detection of Human Phospho-PDGF R beta (Y1021) 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 (Y1021) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2316), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho‑PDGF R beta (Y1021) at approximately 185 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: PDGF R beta
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.
Heldin, C.H. and L. Claesson-Welsh (1994) in Guidebook to Cytokines and Their Receptors, Nicola, N.A. ed. Oxford University Press, New York, p. 202.
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