Cell Signaling Technology

Product Pathways - Lymphocyte Signaling

Btk (D3H5) Rabbit mAb (Alexa Fluor® 488 Conjugate) #82795

Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
F 1:50 Human,Mouse, Endogenous Rabbit IgG

Species cross-reactivity is determined by western blot.

Applications Key: F=Flow Cytometry,

Homology

Species predicted to react based on 100% sequence homology: Rat, Hamster, Bovine, Dog, Pig, Horse,

Specificity / Sensitivity

Btk (D3H5) Rabbit mAb (Alexa Fluor® 488 Conjugate) recognizes endogenous levels of total Btk protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp195 of human Btk protein.

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct flow cytometric analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated Btk (D3H5) Rabbit mAb #8547.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat (blue) and Ramos (green) cells using Btk (D3H5) Rabbit mAb (Alexa Fluor® 488 Conjugate).

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

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

Companion Products


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

Alexa Fluor is a registered trademark of Life Technologies Corporation.

The Alexa Fluor dye conjugates in this product are sold under license from Life Technologies Corporation, for research use only excluding use in combination with DNA microarrays and high content screening (HCS).

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

用户评论 --- 共 0

该产品暂无评论!

我要参与评论 :

如要参与评论请先登录网站

还没有网站账户?去注册一下吧

Products

 

Applications