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558428fig.png

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.

558428fig.png

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.

558428fig.png 558428fig.png Red-Cap-Red-Label.jpg

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.

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.

Product Details
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BD Phosflow™
Mouse (QC Testing), Human (Tested in Development)
Mouse IgG1, κ
Phosphorylated Human PDGFRβ Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_647117
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.
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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. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. 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.
  7. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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558428 Rev. 4
Antibody Details
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J24-618

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.

558428 Rev. 4
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 670-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.
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Alexa Fluor™ 647
653 nm
669 nm
558428 Rev.4
Citations & References
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View product citations for antibody "558428" on CiteAb

Development References (4)

  1. 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).
  2. 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).
  3. Claesson-Welsh L. Platelet-derived growth factor receptor signals. J Biol Chem. 1994; 269(51):32023-32026. (Biology).
  4. 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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558428 Rev. 4

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

 

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.