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Purified Mouse anti-Btk (pY551)/Itk (pY511)
Purified Mouse anti-Btk (pY551)/Itk (pY511)

Western blot analysis of Btk (pY551) in human Burkitt's lymphoma.  Cell lysates from control (Panel A) and pervanadate-treated (Panel B) Raji cells (ATCC CCL-86) were probed with Purified Mouse anti-Btk (pY551)/Itk (pY511) monoclonal antibody at concentrations of 0. 5, 0.25, and 0.125 µg/ml.  Btk (pY551) is identified as a band of about 77 kDa in the treated cell lysate.

Western blot analysis of Btk (pY551) in human Burkitt's lymphoma.  Cell lysates from control (Panel A) and pervanadate-treated (Panel B) Raji cells (ATCC CCL-86) were probed with Purified Mouse anti-Btk (pY551)/Itk (pY511) monoclonal antibody at concentrations of 0. 5, 0.25, and 0.125 µg/ml.  Btk (pY551) is identified as a band of about 77 kDa in the treated cell lysate.

Product Details
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BD Pharmingen™
ATK; BPK; AGMX1
Mouse IgG1, κ
Phosphorylated Human Btk Peptide
Western blot (Routinely Tested)
77 kDa
0.5 mg/ml
AB_2067823
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

Investigators are encouraged to reference http://www.bdbiosciences.com/us/resources/s/cellbiology

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
558034 Rev. 3
Antibody Details
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24a/BTK (Y551)

Bruton's tyrosine kinase (Btk) is a nonreceptor tyrosine kinase whose function is critical for proper B cell development and signaling.  The activity of Btk is regulated by Src mediated phosphorylation of the kinase domain at tyrosine 551 (Y551).  This event induces Btk kinase activity and subsequent autophosphorylation at Y223.  Phosphorylated Btk then associates with the cell membrane via the interaction of the PH domain with phosphatidylinositol 3, 4, 5-triphosphate.  

The Tec family kinase Itk plays a critical role in signal transduction downstream of the T cell antigen receptor and has been implicated in the activation of phospholipase C-γ1 (PLCγ1), a key regulator of calcium mobilization and extracellular signal-regulated kinase (ERK) activation.  Itk is regulated by an activating transphosphorylation event in which Y511 in the kinase domain is phosphorylated by Lck.  

The 24a/BTK (Y551) monoclonal antibody recognizes the Y551-phosphorylated form of human Btk and the Y511 phosphorylated form of human Itk.  

558034 Rev. 3
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
558034 Rev.3
Citations & References
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Development References (3)

  1. Mahajan S, Fargnoli J, Burkhardt AL, Kut SA, Saouaf SJ, Bolen JB. Src family protein tyrosine kinases induce autoactivation of Bruton's tyrosine kinase. Mol Cell Biol. 1995; 15:5304-5311. (Biology). View Reference
  2. Rawlings DJ, Scharenberg AM, Park H, et al. Activation of BTK by a phosphorylation mechanism initiated by SRC family kinases. Science. 1996; 271:822-825. (Biology). View Reference
  3. Wilcox HM, Berg LJ.. Itk phosphorylation sites are required for functional activity in primary T cells. J Biol Chem. 2003; 278:37112-37121. (Biology).
558034 Rev. 3

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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.