Detects human PDGF R alpha in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant human PDGF R beta or recombinant mouse PDGF R alpha is observed.
Monoclonal Mouse IgG1 Clone # 35248
Protein A or G purified from hybridoma culture supernatant
S. frugiperda insect ovarian cell line Sf 21-derived recombinant human PDGF R alpha
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
<0.10 EU per 1 μg of the antibody by the LAL method.
Recombinant Human PDGF R alpha (Catalog # 322-PR) under non-reducing conditions only
Measured by its ability to neutralize PDGF‑AA-induced proliferation in the WS‑1 human fetal skin fibroblast cell line. The Neutralization Dose (ND50) is typically 2-10 µg/mL in the presence of 10 ng/mL Recombinant Human PDGF‑AA.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Cell Proliferation Induced by PDGF‑AA and Neutralization by Human PDGF R alpha Antibody.
Recombinant Human PDGF‑AA (Catalog # 221-AA) stimulates proliferation in the WS‑1 human fetal skin fibroblast cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human PDGF‑AA (10 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human PDGF R alpha Monoclonal Antibody (Catalog # MAB322). The ND50 is typically 2‑10 µg/mL.
PDGF R alpha was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Mouse Anti-Human PDGF R alpha Monoclonal Antibody (Catalog # MAB322) at 25 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
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 alpha
PDGF is a major serum mitogen that can exist as a homo- or heterodimeric 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. Whereas PDGF R alpha binds all three PDGF isoforms with high affinity, PDGF R beta binds PDGF‑BB and AB, but not PDGF-AA. 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 heterodimerization and signal transduction. The expression of the alpha and beta receptors is independently regulated in various cell types. Only PDGF R alpha is expressed in oligodendrocyte progenitor cells, mesothelial cell and liver endothelial cells. Soluble PDGF-R alpha has been detected in cell conditioned medium and human plasma. Recombinant soluble PDGF R alpha binds PDGF with high affinity and is a potent PDGF antagonist (1).
Heldin, C.H. and L. Claesson-Welsh (1994) Guidebook to Cytokines and Their Receptors, Nicola, N.A. (ed) Oxford University Press, New York, NY p. 202.
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