Biolegend, 601702, Purified anti-BTK Phospho (Tyr 223) Antibody, 100μg

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.
100μg
Verified Reactivity: Human, Mouse
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: Human BTK peptide phosphorylated at Tyr 223, Complete Freund's adjuvant.
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 1.0 - 2.0 µg per ml (1:500-1:250). For flow cytometric staining, the suggested use of this reagent is 0.5 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes: A16128B was generated in the same immunization that produced clone A16128C (anti-BTK Phospho (Tyr223)/ITK Phospho (Tyr180)). Both clones bind BTK Phospho Tyr223 with similar affinity, but in comparison to clone A16128C, clone A16128B has weaker recognition of phosphorylated Tyrosine 180 of ITK by intracellular staining and flow cytometric analysis.
RRID: AB_2715906 (BioLegend Cat. No. 601701) AB_2715907 (BioLegend Cat. No. 601702)
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).
Distribution: Cytosol, plasma-membrane associated
Function: Links B 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: Krysiak K, et al. 2017. Blood. 129:473. Burger JA, et al. 2016. Nat Commun. 7:11589. Wang Q, et al. 2015. Elife. 4:e06074. Ito M, et al. 2015. Nat. Commun. 6:7360. Pillinger G, et al. 2015. Sci. Rep. 5:12949. Cinar M, et al. 2013. Leuk. Res. 37:1271. Doyle SL, et al. 2007 J. Biol. Chem. 282:36953. Nore BF, et al. 2003. Biochem. Biophys. Acta. 1645:123. Li Z, et al. 1997. Proc. Natl. Acad. Sci. 94:13820.
Gene ID: 695
UniProt: View information about BTK Phospho Tyr223 on UniProt.org
Clone: A16128B
Regulatory Status: RUO
Other Names: Bruton’s tyrosine kinase, Bruton agammaglobulinemia tyrosine kinase, AGMX1, ATK, IMD1, PSCTK1, XLA, BPK
Isotype: Mouse IgG2a, κ