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Alexa Fluor® 647 Mouse anti-PDGFRβ (CD140b) (pY1009)
Alexa Fluor® 647 Mouse anti-PDGFRβ (CD140b) (pY1009)

Analysis of PDGFRβ (CD140b) (pY1009). NIH/3T3 cells (Mouse embryo fibroblast cells; ATCC CRL-1658) were serum starved overnight and either stimulated with Platelet-Derived Growth Factor-BB (Cat. No. 354051) at 37˚C for 2 minutes (filled histogram) or unstimulated (open histogram).  The cells were fixed with BD Phosflow™ Fix Buffer I (Cat. No. 557870) at 37˚C for 10 minutes, permeabilized in BD Phosflow™ Perm Buffer III (Cat. No. 558050) on ice for at least 30 minutes, and stained with the Alexa Fluor® 647 mouse anti-PDGFRβ (CD140b) (pY1009) antibody.  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry instrument.

Analysis of PDGFRβ (CD140b) (pY1009). NIH/3T3 cells (Mouse embryo fibroblast cells; ATCC CRL-1658) were serum starved overnight and either stimulated with Platelet-Derived Growth Factor-BB (Cat. No. 354051) at 37˚C for 2 minutes (filled histogram) or unstimulated (open histogram).  The cells were fixed with BD Phosflow™ Fix Buffer I (Cat. No. 557870) at 37˚C for 10 minutes, permeabilized in BD Phosflow™ Perm Buffer III (Cat. No. 558050) on ice for at least 30 minutes, and stained with the Alexa Fluor® 647 mouse anti-PDGFRβ (CD140b) (pY1009) antibody.  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry instrument.

Product Details
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BD Phosflow™
CD140b (pY1009)
Mouse (QC Testing), Human (Reactivity Confirmed in Development)
Mouse IgG2b, κ
Phosphorylated Human PDGFRβ (pY1009) Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_647114
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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was 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. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. 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.
  5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  6. 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.
  7. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  8. All other brands are trademarks of their respective owners.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
558323 Rev. 2
Antibody Details
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J25-602

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 (PLC-γ).

The J25-602 monoclonal antibody recognizes the phosphorylated tyrosine 1009 (pY1009) in the C-terminal noncatalytic region of CD140b, which interacts primarily with protein tyrosine phosphatase 1D and possibly with PLC-γ. The orthologous phosphorylation site in mouse PDGFRβ is Y1008.

558323 Rev. 2
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
558323 Rev.2
Citations & References
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Development References (2)

  1. Claesson-Welsh L. Platelet-derived growth factor receptor signals. J Biol Chem. 1994; 269(51):32023-32026. (Biology). View Reference
  2. 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
558323 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.