Biolegend, 646901, Purified anti-BTK Phospho (Tyr551)/ITK Phospho (Tyr512) Antibody, 25μg

Bruton’s tyrosine kinase (BTK) is a Tec family kinase that plays a critical role in B cell development. Upon B cell receptor engagement, BTK translocates to the plasma membrane, where it is transphosphorylated by LYN and SYK kinases at Tyr 551. This initial phosphorylation event is followed by autophosphorylation at Tyr 223. Phosphorylated Tyr 223 is proposed to serve as a docking site for other proteins containing an SH2 domain. Downregulation of BTK activity is achieved through phosphorylation at Ser 180 by PKCβ, which results in reduced membrane recruitment and concomitant transphosphorylation. BTK-mediated B cell receptor signaling is required for B cell survival in the bone marrow and is a therapeutic target for certain B cell malignancies. Mutations in the BTK gene results in X-linked agammaglobulinemia (XLA), a primary immunodeficiency disease characterized by a failure in B cell maturation. Additionally, BTK plays a role in Toll-like receptor signaling in innate immune cells, and has been shown to play a direct role in NLRP3 inflammasome activation, thus contributing to ischaemic brain injury. IL2-inducible T-cell kinase (ITK) is a tyrosine kinase that is expressed in T cells. It regulates the proliferation, differentiation and development of conventional T cells and NK T cells. Once T cells are activated, a series of phosphorylation events occur leading to the phosphorylation of Tyr 512 in the activation loop by LCK. Subsequent autophosphorylation at Tyr 180 leads to activation of the kinase. Activated ITK can stimulated downstream intracellular calcium mobilization and NFAT nucleus translocation, and in turn, activate T cells. The phenotypes of mice deficient in ITK show severely affected immune response and development.
25μg
Verified Reactivity: Human, Mouse
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: Human BTK peptide phosphorylated at Tyr 551.
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation: The antibody was purified by affinity chromatography.
Concentration: 0.5 mg/ml
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C.
Application: WB - Quality tested ICFC - Verified
Recommended Usage: Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.125 - 2.0 µg/ml (1:250-4000 dilution). For intracellular flow cytometric staining, the suggested use of this reagent is ≤ 0.25 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes: This clone was initially generated against a BTK peptide sequence containing phospho Tyrosine 551, but was shown to also bind to the highly homologous phospho Tyrosine 512 within the T-cell specific tyrosine kinase ITK. Due to complete conservation of these BTK and ITK phosphosites and the surrounding residues between human and mice, this antibody is predicted to recognize mouse BTK Phospho Tyrosine 551 and mouse ITK Phospho Tyrosine 517.
RRID: AB_2721728 (BioLegend Cat. No. 646901) AB_2721729 (BioLegend Cat. No. 646902)
Structure: BTK encodes a 659 amino acid protein with a predicted molecular weight of 76.2 kD. Contains a PH domain (a.a. 1-133) that binds phosphatidylinositol (3,4,5)-triphosphate (PIP3) and polyphosphates, an SH3 domain (a.a. 214-274), an SH2 domain (a.a. 281-377), and a protein kinase domain (402-655).ITK encodes a 620 amino acid protein with a predicted molecular weight of 71.8 kD. Contains one PH domain (a.a. 4-111) for plasma membrane recruitment, both SH3 (a.a. 171-231) and SH3 (a.a. 239-338) domains, and one protein kinase domain (a.a. 363-615).
Distribution: BTK: Cytosol, plasma membrane-associated, B cells ITK: Cytosol, plasma membrane-associated, T cells
Function: BTK: Links B cell receptor engagement with downstream signaling events ITK: Links T cell receptor engagement with downstream signaling events
Interaction: ARID3A, BLNK, CAV1, GNAQ, GTF2I, PLGC2, PRKD1, SH3BP5
Ligand/Receptor: PIP3
Biology Area: Cell Biology, Signal Transduction
Molecular Family: Phospho-Proteins, Protein Kinases/Phosphatase
Antigen References: Yang W, et al. 1997. Proc Natl Acad Sci U.S.A. 94:604. Park H, et al. 1996. Immunity. 4:515. Lin L, et al. 2009. Biochemistry. 48:2021. Heyeck SD, et al. 1997. J. Biol. Chem. 272:25401. Nore BF, et al. 2003. Biochim. Biophys. Acta. 1645:123.
Gene ID: 6953702
UniProt: View information about BTK Phospho Tyr551/ITK Phospho Tyr512 on UniProt.org
Clone: A16064A
Regulatory Status: RUO
Other Names: BTK: Bruton’s tyrosine kinase, Bruton agammaglobulinemia tyrosine kinase, AGMX1, ATK, IMD1, PSCTK1, XLA, BPK, ITK: Interleukin-2-inducible T-cell kinase, T-cell-specific kinase, EMT, LYK, PSCTK2
Isotype: Mouse IgG1, κ