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PE 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
Ave. Rating
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Product Citations
publications
A16128C_PE_BTK_Phospho_Antibody_1_052319_updated.png
Human peripheral blood lymphocytes were treated with (left), or without (right) Hydrogen Peroxide for 5 minutes, fixed with Fixation Buffer (Cat No. 420801), permeabilized with Intracellular Staining Permeabilization Wash Buffer (Cat No. 421002), then stained with CD19 Brilliant Violet 421™ and anti-BTK Phospho (Tyr223)/ITK Phospho (Tyr180) (Clone A16128C) PE.
  • A16128C_PE_BTK_Phospho_Antibody_1_052319_updated.png
    Human peripheral blood lymphocytes were treated with (left), or without (right) Hydrogen Peroxide for 5 minutes, fixed with Fixation Buffer (Cat No. 420801), permeabilized with Intracellular Staining Permeabilization Wash Buffer (Cat No. 421002), then stained with CD19 Brilliant Violet 421™ and anti-BTK Phospho (Tyr223)/ITK Phospho (Tyr180) (Clone A16128C) PE.
  • A16128C_PE_BTK_Phospho_Antibody_2_052319_updated.png
    C57BL/6 mouse splenocytes were treated with (left), or without (right) hydrogen peroxide for 5 minutes, fixed with Fixation Buffer (Cat. No. 420801), permeabilized with Intracellular Staining Permeabilization Wash Buffer (Cat. No. 421002), then stained with CD19 APC and anti-BTK Phospho (Tyr223)/ITK Phospho (Tyr180) (Clone A16128C) PE.
See PE spectral data
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605203 25 tests 159 CHF
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605204 100 tests 360 CHF
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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 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 (pdf)

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 and BSA (origin USA)
Preparation
The antibody was purified by affinity chromatography and conjugated with PE under optimal conditions.
Concentration
Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

ICFC - Quality tested
Recommended Usage

Each lot of this antibody is quality control tested by intracellular flow cytometry using our True-Phos™ Perm Buffer in Cell Suspensions Protocol. For flow cytometric staining, the suggested use of this reagent is 5 µl per million cells in 100 µl staining volume or 5 µl per 100 µl of whole blood.

Excitation Laser
Blue Laser (488 nm)
Green Laser (532 nm)/Yellow-Green Laser (561 nm)
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_2721622 (BioLegend Cat. No. 605203)
AB_2721622 (BioLegend Cat. No. 605204)

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

Related FAQs

What type of PE do you use in your conjugates?
We use R-PE in our conjugates.
Go To Top Version: 0    Revision Date: 01.05.2018

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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