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Alexa Fluor® 647 Mouse Anti–c-Cbl (pY774)

Alexa Fluor® 647 Mouse Anti–c-Cbl (pY774)

クローン 29/c-Cbl

(RUO)
Alexa Fluor® 647 Mouse Anti–c-Cbl (pY774)
Analysis of c-Cbl (pY774) in activated human T leukemia cells. Jurkat cells (ATCC TIB152) were serum starved overnight and then either stimulated with 5 mM hydrogen peroxide 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 (pY774).  Flow cytometry was performed on a BD™ FACSCalibur flow cytometry system.
Analysis of c-Cbl (pY774) in activated human T leukemia cells. Jurkat cells (ATCC TIB152) were serum starved overnight and then either stimulated with 5 mM hydrogen peroxide 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 (pY774).  Flow cytometry was performed on a BD™ FACSCalibur flow cytometry system.
製品詳細
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BD Phosflow™
Human (QC Testing)
Mouse IgG1, κ
Phosphorylated Human c-Cbl Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_397028
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 antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  6. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  7. 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.
  8. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  9. All other brands are trademarks of their respective owners.
558103 Rev. 2
抗体の詳細
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29/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, 731, and 774 (Y774) 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 Y774 site interacts with the SH2 domain of the CRK adapter protein.  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 29/c-Cbl monoclonal antibody recognizes the Y774-phosphorylated form of human c-Cbl.

558103 Rev. 2
引用&参考文献
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Development References (2)

  1. Thien CBF, Langdon WY. CBL: Many adaptations to regulate protein tyrosine kinase. Nat Rev Mol Cell Biol. 2001; 2:294-307. (Biology).
  2. Tsygankov AY, Teckchandani AM, Feshchenko EA, Swaminathan G. Beyond the RING: CBL proteins as multivalent adapters. Oncogene. 2001; 20:6382-6402. (Biology).
558103 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.