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PE Mouse anti-PDGFRβ (CD140b) (pY771)
PE Mouse anti-PDGFRβ (CD140b) (pY771)

Analysis of PDGFRβ (CD140b) (pY771) 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 PE anti-PDGFRβ (CD140b) (pY771, Cat. No. 558426).  Flow cytometry was performed on a

BD FACSCalibur™ flow cytometry system.

Analysis of PDGFRβ (CD140b) (pY771) 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 PE anti-PDGFRβ (CD140b) (pY771, Cat. No. 558426).  Flow cytometry was performed on a

BD FACSCalibur™ flow cytometry system.

Product Details
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BD Phosflow™
Mouse (QC Testing), Human (Predicted)
Mouse IgG1, κ
Phosphorylated Human PDGFRβ Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_647189
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Product Notices

  1. 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).
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. 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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
558426 Rev. 2
Antibody Details
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J23-1044

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 J23-1044 monoclonal antibody recognizes the phosphorylated tyrosine 771 (pY771) in the kinase insert domain of CD140b.  pY771 interacts with GTPase-activating protein, a negative regulator of Ras, and weakly with Shc, which indirectly promotes the activation of Ras.

558426 Rev. 2
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
558426 Rev.2
Citations & References
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Development References (3)

  1. Claesson-Welsh L. Platelet-derived growth factor receptor signals. J Biol Chem. 1994; 269(51):32023-32026. (Biology).
  2. Ekman S, Kallin A, Engstrom U, Heldin CH, Ronnstrand L. SHP-2 is involved in heterdimer specific loss of phosphorylation of Tyr771 in the PDGF beta-receptor. Oncogene. 2002; 21(12):1870-1875. (Biology).
  3. 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
558426 Rev. 2

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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.