antibody

Btk, phosphorylated (Ser180) (Bruton’s Tyrosine Kinase)

B5000-10

100 ul

monoclonal

mouse

nonconjugated

phosphorylated

3i5(3D3)

Btk, phosphorylated (Ser180) (Bruton’s Tyrosine Kinase)

mouse

Mab

Phosphorylated

Antibodies

100 ul

3i5(3D3)

IgG1

Bruton’s tyrosine kinase (Btk) is a member of the Btk/Tec family of cytoplasmic tyrosine kinases. Like other Btk family members, it contains a pleckstrin homology (PH) domain, Src homology SH3 and SH2 domains. Btk plays an important role in B cell development. Activation of B cells by various ligands is accompanied by Btk membrane translocation mediated by its PH domain binding to phosphatidylinositol-3,4,5-trisphosphate. The membrane-located Btk is active and associated with transient phosphorylation of two tyrosine residues, Tyr551 and Tyr223. Tyr551 in the activation loop is transphosphorylated by the Src family tyrosine kinase, leading to autophosphorylation at Tyr223 within the SH3 domain, which is necessary for full activation. The activation of Btk is negatively regulated by PKCbeta through phosphorylation of Btk at Ser180, which results in reduced membrane recruitment, transphosphorylation and subsequent activation. The PKC/Btk inhibitory signal is likely to be a key determinant of the B-cell receptor signaling threshold to maintain optimal Btk activity. Applications: Suitable for use in ELISA and Western Blot. Other applications not tested. Recommended Dilutions: Western Blot: 1:1000; For Western Blot, USBio recommends using immunoprecipitated Btk. Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Hu

Synthetic phosphopeptide corresponding to residues around Ser180 of human Btk. Species Sequence Homology: mouse, 100%.

Recognizes transfected levels of human Btk only when phosphorylated at Ser180 at ~80kD.

Ascites

Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol.

Not Determined

Product Type: Mab Isotype: IgG1 Clone No: 3i5(3D3) Host: mouse Source: human Concentration: Not Determined Form: Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. Purity: Ascites Immunogen: Synthetic phosphopeptide corresponding to residues around Ser180 of human Btk. Species Sequence Homology: mouse, 100%. Specificity: Recognizes transfected levels of human Btk only when phosphorylated at Ser180 at ~80kD. Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

E WB

-20°C

1. Khan, W.N., Immunol. Res. 23: 147-156 (2001). 2. Lewis, C.M., et al., Curr. Opin. Immunol. 13: 317-325 (2001). 3. Salim, K., et al., EMBO J. 15: 6241–6250 (1996). 4. Rameh, L.E., et al., J. Biol. Chem. 272: 22,059–22,066 (1997). 5. Varnai, P., et al., J. Biol. Chem. 274: 10,983–10,989 (1999). 6. Rawlings, D.J., et al., Science 271: 822–825 (1996). 7. Park, H., et al., Immunity 4: 515–525 (1996). 8. Kang, S.W., et al., EMBO J. 20: 5692–5702 (2001).

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