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Analysis of PDGFRβ (CD140b) (pY857) in mouse embryonic fibroblasts. Serum-starved NIH/3T3 cells were either stimulated with PDGF-BB (shaded histogram) or unstimulated (open histogram). The cells were fixed (BD Phosflow™ Fix Buffer I, Cat. No. 557870) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with Alexa Fluor® 647 Mouse anti-PDGFRβ (CD140b) (pY857, Cat. No. 558428). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
BD™ Phosflow Alexa Fluor® 647 Mouse anti-PDGFRβ (CD140b) (pY857)
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Platelet-derived growth factor (PDGF) is a potent mitogen for cells of mesenchymal origin and exerts its effects by binding to the PDGF receptor (PDGFR), a transmembrane protein tyrosine kinase. PDGFR is composed of PDGFRα (CD140a) and/or PDGFRβ (CD140b) polypeptides. Both PDGF and PDGFR consist of subunits that form homo- or heterodimers with varying specificities: PDGF-AA binds only to αα PDGFR, PDGF-AB binds to both αα and αβ PDGFR, and PDGF-BB binds to all three PDGFRs. Ligand binding induces dimerization and activation of the receptor. Upon activation, CD140b is phosphorylated at multiple tyrosine sites and, in turn, an intracellular phosphorylation cascade is initiated. PDGFR localizes primarily to membrane invaginations termed caveolae, compartments that are enriched in several of its downstream effectors, including phosphatidylinositol 3'-kinase, Src, and phospholipase C-γ.
The J24-618 monoclonal antibody recognizes the phosphorylated tyrosine 857 (pY857) in the tyrosine kinase domain 2 of CD140b, which is required for maximal receptor kinase activity. The orthologous phosphorylation site in mouse PDGFRβ is Y856.
Development References (4)
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Baxter RM, Secrist JP, Vaillancourt RR, Kazlauskas A. Full activation of the platelet-derived growth factor β-receptor kinase involves multiple events. J Biol Chem. 1998; 273(27):17050-17055. (Biology).
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Chiarugi P, Cirri P, Taddei ML, Giannoni E, et al. Insight into the role of low molecular weight phosphotyrosin phosphatase (LMW-PTP) on platelet-derived growth factor receptor (PDGF-r) signaling. J Biol Chem. 2002; 277(40):37331-37338. (Biology).
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Claesson-Welsh L. Platelet-derived growth factor receptor signals. J Biol Chem. 1994; 269(51):32023-32026. (Biology).
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Liu J, Oh P, Horner T, Rogers RA, Schnitzer JE. Organized endothelial cell surface signal transduction in caveolae distinct from glycosylphosphatidylinositol-anchored protein microdomains. J Biol Chem. 1997; 272(11):7211-7222. (Biology). View Reference
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