Btk (pY223)/Itk (pY180) Mouse, Brilliant Violet 421, Clone: N35-86, BD, Mouse Monoclonal Antibody, Each

Details

The N35-86 reacts with human and mouse Bruton's tyrosine kinase that is phosphorylated at the tyrosine 223 position, Btk (pY223). Btk is also known as agammaglobulinaemia tyrosine kinase (ATK) and B-cell progenitor kinase (BPK). Btk is a nonreceptor tyrosine kinase whose function is critical for proper B cell development and signaling. It is a member of the Tec family of kinases which includes Tec and Itk. In addition to an N-terminal pleckstrin homology (PH) domain, the Tec proteins contain Src homology domains 2 and 3 (SH2 and SH3) and a stretch of 60-80 amino acids between the PH and SH3 domains termed the Tec homology domain. The activity of Btk is regulated by Src-mediated phosphorylation of the kinase domain at tyrosine 551. This event induces Btk kinase activity and subsequent autophosphorylation at tyrosine 223 in the SH3 domain. Phosphorylated Btk then associates with the cell membrane via the interaction of the PH domain with phosphatidylinositol 3, 4, 5-triphosphate. The PH domain is essential for proper activation and function of Btk. A mutation in the PH domain results in Xid, murine X-linked immunodeficiency, and human X-linked agammaglobulinemia. The orthologous phosphorylation site for rat BTK is Y224. Crossreactivity with human Itk (pY180) was confirmed by immunoprecipitation and Western blot analyses using the N35-86 antibody. The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.