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Alexa Fluor® 647 Mouse anti-c-Cbl (pY700)
Alexa Fluor® 647 Mouse anti-c-Cbl (pY700)

Analysis of c-Cbl (pY700) in activated human T leukemia cells.  Jurkat cells (ATCC TIB152) were serum starved overnight and then either stimulated with 5 mM hydrogen peroxide (H2O2) for 15 minutes (shaded histogram) or unstimulated (open histogram).  The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes, then permeabilized (BD PhosFlow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with Alexa Fluor® 647 anti-c-Cbl (pY700).  Flow cytometry was performed on a

BD FACSCalibur™ flow cytometry system.

Analysis of c-Cbl (pY700) in activated human T leukemia cells.  Jurkat cells (ATCC TIB152) were serum starved overnight and then either stimulated with 5 mM hydrogen peroxide (H2O2) for 15 minutes (shaded histogram) or unstimulated (open histogram).  The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes, then permeabilized (BD PhosFlow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with Alexa Fluor® 647 anti-c-Cbl (pY700).  Flow cytometry was performed on a

BD FACSCalibur™ flow cytometry system.

商品详情
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BD Phosflow™
Human (QC Testing), Mouse (Tested in Development)
Mouse IgG1
Human c-Cbl (pY700) Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


准备和存储

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.

商品通知

  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  3. 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.
  4. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  5. 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.
  6. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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.
558100 Rev. 3
抗体详情
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47/c-Cbl

Cbl (Casitas B-lineage lymphoma) was identified in the genome of a transforming retrovirus from a mouse pre-B lymphoma.  The cellular gene product c-Cbl is one of numerous Cbl-related proteins found in vertebrate and invertebrate organisms.  It is an 120-kDa adapter protein that contains multiple functional domains, including a RING finger motif, a tyrosine kinase-binding (TKB) domain, and a proline-rich region.  The TKB domain directly interacts with specific auto-phosphorylation sites in activated protein-tyrosine kinases (PTK).  Through the RING finger motif, c-Cbl recruits and activates an E2 ubiquitin-conjugating enzyme, thus targeting the activated PTK for protein degradation.  The proline-rich region contains SH3 domain-binding and 14-3-3 protein-binding motifs.  c-Cbl is also phosphorylated at tyrosines 700 (Y700), 731, and 774 by Syk- and Src-family kinases after the stimulation of some integrins and a wide variety of receptors for antigens, immunoglobulins, growth factors, cytokines, and hormones.  In turn, the phosphorylated Y700 site interacts with the SH2 domains of CRK and Vav1.  The c-Cbl adapter protein is expressed in the cytoplasm in all tissues, with especially high levels of expression in hematopoietic cells.  Through its many functional sites, c-Cbl plays key roles in the positive and negative regulation of vital cell functions, including T Cell Receptor-mediated cellular immune responses.

The 47/c-Cbl monoclonal antibody recognizes the Y700-phosphorylated form of human c-Cbl.  The orthologous phosphorylation site in mouse c-Cbl is Y698.

558100 Rev. 3
格式详情
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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
558100 Rev.3
报价单和参考
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研发参考 (3)

  1. Miura-Shimura Y, Duan L, Rao NL, et al. Cbl-mediated ubiquitinylation and negative regulation of Vav. J Biol Chem. 2003; 278:38495-38504. (Biology).
  2. Thien CBF, Langdon WY. CBL: Many adaptations to regulate protein tyrosine kinases. Nat Rev Mol Cell Biol. 2001; 2:294-307. (Biology).
  3. Tsygankov AY, Teckchandani AM, Feshchenko EA, Swaminathan G. Beyond the RING: CBL proteins as multivalent adapters. Oncogene. 2001; 20:6382-6402. (Biology).
558100 Rev. 3

Please refer to Support Documents for Quality Certificates


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.