Phospho-Btk (Tyr551) (E5Y6N) Rabbit mAb #18805
- WB
- IP
- F
Supporting Data
| REACTIVITY | H |
| SENSITIVITY | Endogenous |
| MW (kDa) | 78 |
| Source/Isotype | Rabbit IgG |
Application Key:
- WB-Western Blotting
- IP-Immunoprecipitation
- F-Flow Cytometry
Species Cross-Reactivity Key:
- H-Human
Product Information
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:50 |
| Flow Cytometry (Fixed/Permeabilized) | 1:200 - 1:800 |
Storage
For a carrier free (BSA and azide free) version of this product see product #69360.
Protocol
Specificity / Sensitivity
Phospho-Btk (Tyr551) (E5Y6N) Rabbit mAb recognizes endogenous levels of Btk protein only when phosphorylated at Tyr551.
Species Reactivity:
Human
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr551 of human Btk protein.
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).
- Khan, W.N. (2001) Immunol Res 23, 147-56.
- Lewis, C.M. et al. (2001) Curr Opin Immunol 13, 317-25.
- Salim, K. et al. (1996) EMBO J 15, 6241-50.
- Rameh, L.E. et al. (1997) J Biol Chem 272, 22059-66.
- Várnai, P. et al. (1999) J Biol Chem 274, 10983-9.
- Rawlings, D.J. et al. (1996) Science 271, 822-5.
- Park, H. et al. (1996) Immunity 4, 515-25.
- Kang, S.W. et al. (2001) EMBO J 20, 5692-702.
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