Purified anti-BTK Phospho (Tyr 223) Antibody

Pricing & Availability
Clone
A16128B (See other available formats)
Other Names
Bruton’s tyrosine kinase, Bruton agammaglobulinemia tyrosine kinase, AGMX1, ATK, IMD1, PSCTK1, XLA, BPK
Isotype
Mouse IgG2a, κ
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Product Citations
publications
1_A16128B_PURE_BTKphospho_WB_081517
Total cell lysates (15 µg protein) from Ramos cells treated with (+) or without (-) 5 mM H2O2 for 3 minutes were resolved by 4-20% Tris-Glycine gel electrophoresis, transferred to nitrocellulose, and probed with 2.0 µg/mL (1:250) purified anti-BTK Phospho (Tyr223) antibody, clone A16128B. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse-IgG (Cat. No. 405301) at a 1:3000 dilution. Equal BTK loading was confirmed using a pan-BTK antibody.
  • 1_A16128B_PURE_BTKphospho_WB_081517
    Total cell lysates (15 µg protein) from Ramos cells treated with (+) or without (-) 5 mM H2O2 for 3 minutes were resolved by 4-20% Tris-Glycine gel electrophoresis, transferred to nitrocellulose, and probed with 2.0 µg/mL (1:250) purified anti-BTK Phospho (Tyr223) antibody, clone A16128B. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse-IgG (Cat. No. 405301) at a 1:3000 dilution. Equal BTK loading was confirmed using a pan-BTK antibody.
  • A16128B_PURE_BTK_Phospho_Antibody_2_052319_updated.png
    Human peripheral blood lymphocytes were treated with (left), or without (right) hydrogen peroxide for 5 minutes, fixed with Fixation Buffer, permeabilized with Intracellular Staining Permeabilization Wash Buffer, then stained with CD20 (clone 2H7) APC and purified anti-BTK Phospho (Tyr223) (Clone A16128B). Followed by anti-mouse IgG2a, κ PE.
Cat # Size Price Quantity Avail. Save
601701 25 µg $85
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601702 100 µg $215
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Description

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.

Product Details
Technical data sheet

Product Details

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 - Validated

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)

Antigen Details

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
  1. Krysiak K, et al. 2017. Blood. 129:473.
  2. Burger JA, et al. 2016. Nat Commun. 7:11589.
  3. Wang Q, et al. 2015. Elife. 4:e06074.
  4. Ito M, et al. 2015. Nat. Commun. 6:7360.
  5. Pillinger G, et al. 2015. Sci. Rep. 5:12949.
  6. Cinar M, et al. 2013. Leuk. Res. 37:1271.
  7. Doyle SL, et al. 2007 J. Biol. Chem. 282:36953.
  8. Nore BF, et al. 2003. Biochem. Biophys. Acta. 1645:123.
  9. Li Z, et al. 1997. Proc. Natl. Acad. Sci. 94:13820.
Gene ID
695 View all products for this Gene ID
UniProt
View information about BTK Phospho (Tyr223) on UniProt.org

Related FAQs

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Go To Top Version: 2    Revision Date: 05/04/2018

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