Detection of Human and Mouse Phospho-PDGF R beta (Y751) by Western Blot.
Western blot shows lysates of CCD‑1070Sk human foreskin fibroblast cell line and NIH‑3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human PDGF‑BB (Catalog # 220-BB) for 20 minutes. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse Phospho-PDGF R beta (Y751) Monoclonal Antibody (Catalog # MAB9027) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). 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.
Phospho-PDGF R beta (Y751) in BJ Human Cell Line.
PDGF R beta phosphorylated at Y751 was detected in immersion fixed BJ human skin fibroblast cell line stimulated (top panel), and unstimulated (bottom panel), with Recombinant Human PDGF-BB (Catalog # 220-BB) using Rabbit Anti-Human/Mouse Phospho-PDGF R beta (Y751) Monoclonal Antibody (Catalog # MAB9027) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Mouse Phospho-PDGF R beta (Y751) by Simple WesternTM.
Simple Western lane view shows lysates of NIH‑3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human PDGF‑BB (Catalog # 220-BB) for 20 minutes, loaded at 0.2 mg/mL. A specific band was detected for PDGF R beta at approximately 240 kDa (as indicated) using 1 µg/mL of Rabbit Anti-Human/Mouse Phospho-PDGF R beta (Y751) Monoclonal Antibody (Catalog # MAB9027). This experiment was conducted under reducing conditions and using the 66-440 kDa separation system.
Preparation and Storage
Reconstitute at 0.5 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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