Cell Signaling Technology

Product Pathways - Lymphocyte Signaling

Btk (D6T2C) Mouse mAb #56044

No. Size Price
56044S 100 µl ( 10 western blots ) ¥3,250.00 现货查询 购买询价 防伪查询
56044 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse, Endogenous 78 Mouse IgG2b
IP 1:200
F 1:800

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, F=Flow Cytometry,

Specificity / Sensitivity

BTK (D6T2C) Mouse mAb recognizes endogenous levels of total Btk protein. The antibody is predicted to recognize two known Btk isoforms (Btk-A and Btk-C), which are derived from the same gene, but regulated by alternative promoter usage.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human Btk protein. The region is 100% conserved between Btk-A and Btk-C isoforms.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells (blue) and Ramos cells (green) using BTK (D6T2C) Mouse mAb. Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Btk (D6T2C) Mouse mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Extracts from T cells (Jurkat and MOLT4) are negative for Btk expression, demonstrating specificity of the antibody.

IP

IP

Immunoprecipitation of Btk from Ramos cell extracts. Lane 1 is 10% input, lane 2 is Mouse (G3A1) mAb IgG1 Isotype Control #5415, and lane 3 is Btk (D6T2C) Mouse mAb. Western blot analysis was performed with Btk (D6T2C) Rabbit mAb #8547.

Background

Bruton's tyrosine kinase (Btk) is a member of the Btk/Tec family of cytoplasmic tyrosine kinases. Like other Btk family members, it contains a pleckstrin homology (PH) domain and Src homology SH3 and SH2 domains. Btk plays an important role in B cell development (1,2). Activation of B cells by various ligands is accompanied by Btk membrane translocation mediated by its PH domain binding to phosphatidylinositol-3,4,5-trisphosphate (3-5). The membrane-localized Btk is active and associated with transient phosphorylation of two tyrosine residues, Tyr551 and Tyr223. Tyr551 in the activation loop is transphosphorylated by the Src family tyrosine kinases, leading to autophosphorylation at Tyr223 within the SH3 domain, which is necessary for full activation (6,7). The activation of Btk is negatively regulated by PKCβ through phosphorylation of Btk at Ser180, which results in reduced membrane recruitment, transphosphorylation, and subsequent activation (8). The PKC inhibitory signal is likely to be a key determinant of the B cell receptor signaling threshold to maintain optimal Btk activity (8).

  1. Khan, W.N. (2001) Immunol Res 23, 147-56.
  2. Lewis, C.M. et al. (2001) Curr Opin Immunol 13, 317-25.
  3. Salim, K. et al. (1996) EMBO J 15, 6241-50.
  4. Rameh, L.E. et al. (1997) J Biol Chem 272, 22059-66.
  5. Várnai, P. et al. (1999) J Biol Chem 274, 10983-9.
  6. Rawlings, D.J. et al. (1996) Science 271, 822-5.
  7. Park, H. et al. (1996) Immunity 4, 515-25.
  8. Kang, S.W. et al. (2001) EMBO J 20, 5692-702.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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