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Purified anti-BTK Phospho (Tyr223)/ITK Phospho (Tyr180) Antibody

Pricing & Availability
Clone
A16128C (See other available formats)
Regulatory Status
RUO
Other Names
Bruton’s tyrosine kinase, Bruton agammaglobulinemia tyrosine kinase, AGMX1, ATK, IMD1, PSCTK1, XLA, BPK
Isotype
Mouse IgG1, κ
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Product Citations
publications
1_A16128C_PURE_BTKITK_Phospho_Antibody_WB_101217
Total cell lysates (15 µg protein) from Ramos cells treated without (lane 1) or with (lane 2) 2 mM H2O2 for three minutes, and Jurkat cells treated without (lane 3) or with (lane 4) the same stimulation were resolved by 4-20% Tris-Glycine gel electrophoresis, transferred to nitrocellulose, and probed with 2 µg/mL (1:250) purified anti-BTK/ITK Phospho (Tyr223/Tyr180) antibody (Clone A16128C). Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse-IgG. Equal BTK and ITK loading were confirmed using pan antibodies (1 µg/mL, 1:500 dilution) against the respective proteins.
  • 1_A16128C_PURE_BTKITK_Phospho_Antibody_WB_101217
    Total cell lysates (15 µg protein) from Ramos cells treated without (lane 1) or with (lane 2) 2 mM H2O2 for three minutes, and Jurkat cells treated without (lane 3) or with (lane 4) the same stimulation were resolved by 4-20% Tris-Glycine gel electrophoresis, transferred to nitrocellulose, and probed with 2 µg/mL (1:250) purified anti-BTK/ITK Phospho (Tyr223/Tyr180) antibody (Clone A16128C). Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse-IgG. Equal BTK and ITK loading were confirmed using pan antibodies (1 µg/mL, 1:500 dilution) against the respective proteins.
  • A16128C_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)/ITK Phospho (Tyr180) (Clone A16128C). Followed by anti-mouse IgG1, κ PE.
Cat # Size Price Save
605201 25 µg ¥24,640
605202 100 µg ¥58,300
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 Tyr551. This initial phosphorylation event is followed by autophosphorylation at Tyr223. Phosphorylated Tyr223 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

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 2.0 ug/mL (1:250 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

Clones A16128C and A16128B (anti-BTK Phospho (Tyr223)) both bind BTK Phospho Tyr223 with similar affinity, but in comparison to clone A16128B, clone A16128C has a much stronger recognition of phosphorylated Tyrosine 180 of ITK by intracellular staining and flow cytometric analysis. ITK is the T cell-specific homolog of BTK that is not found in B cells. Similarly, T cells lack BTK1. 

Application References

(PubMed link indicates BioLegend citation)
  1. Tomlinson MG, et al. 2004. Mol. Cell. Biol. 24: 2455
RRID
AB_2721726 (BioLegend Cat. No. 605201)
AB_2721726 (BioLegend Cat. No. 605202)

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, B cells

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
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/ITK Phospho Tyr180 on UniProt.org
Go To Top Version: 0    Revision Date: 10/13/2017

For Research Use Only. Not for diagnostic or therapeutic use.

 

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Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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