CST, 8547S, Btk (D3H5) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying BTK. Validated for WB,WB,IP,IHC,IHC,IF,F. Available in 3 sizes. Highly specific and rigorously validated in-house, Btk (D3H5) Rabbit Monoclonal Antibody (CST #8547) is ready to ship. Product Usage Information Western Blotting: 1:1000 Simple Western™: 1:10 - 1:50 Immunoprecipitation: 1:200 IHC Leica Bond: 1:50 - 1:200 Immunohistochemistry (Paraffin): 1:50 - 1:200 Immunofluorescence (Immunocytochemistry): 1:200 - 1:800 Flow Cytometry (Fixed/Permeabilized): 1:100 - 1:400 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody. For a carrier free (BSA and azide free) version of this product see product # 94988 . Protocol Available protocols: Western Blotting, Immunoprecipitation, IHC Leica Bond, Immunohistochemistry (Paraffin), Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized) Specificity / Sensitivity Btk (D3H5) Rabbit Monoclonal Antibody recognizes endogenous levels of total Btk protein. Species Reactivity: Human, Mouse Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp195 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). Alternate Names Agammaglobulinaemia tyrosine kinase; Agammaglobulinemia tyrosine kinase; AGMX1; AT; ATK; B-cell progenitor kinase; BPK; Bruton agammaglobulinemia tyrosine kinase; Bruton tyrosine kinase; Bruton's tyrosine kinase; BTK; dominant-negative kinase-deficient Brutons tyrosine kinase; IGHD3; IMD1; MGC126261; MGC126262; PSCTK1; truncated Bruton agammaglobulinemia tyrosine kinase; Tyrosine-protein kinase BTK; XLA Specification REACTIVITY: H M SENSITIVITY: Endogenous MW (kDa): 77 Source/Isotype: Rabbit IgG