Product Pathways - Lymphocyte Signaling
Blk Kinase #7444
Cell Signaling Technology offers a full line of protein kinases, substrates, antibody detection reagents and HTScan® kits. Browse our "Reagents for High-Throughput Screening" product listing or contact us at drugdiscovery@cellsignal.com.
Description
Purified recombinant full length human Blk (Met1-Pro505) kinase, supplied as a GST fusion protein.
Source / Purification
The GST-Kinase fusion protein was produced using a baculovirus expression system with a construct expressing human Blk (Met1-Pro505) (GenBank Accession No. BC007371) with an amino-terminal GST tag. The protein was purified by one-step affinity chromatography using glutathione-agarose.
Gel Staining
Figure 1. The purity of the GST-Blk fusion protein was analyzed using SDS/PAGE followed by Coomassie stain.
Kinase Assay - Radiometric
Figure 2. Blk kinase activity was measured in a radiometric assay using the following reaction conditions: 4 mM MOPS, pH 7.2, 2.5 mM β-glycerophosphate, 1 mM EGTA, 0.4 mM EDTA, 10 mM MnCl2, 4 mM MgCl2, 0.05 mM DTT, 50 μM ATP, Substrate: Poly(Glu-Tyr), 400 ng/μL and recombinant Blk: variable.
Quality Control
The theoretical molecular weight of the GST-Blk fusion protein is 84 kDa. The purified kinase was quality controlled for purity using SDS-PAGE followed by Coomassie stain [Fig.1]. Blk kinase activity was determined using a radiometric assay [Fig.2].
Background
Blk is a Src family protein tyrosine kinase expressed in all stages of B cell development (1,2). Activation of B cells by various ligands is accompanied by activation of Blk (3). It has been suggested that Blk is involved in the control of B cell differentiation and proliferation (4,5). Blk transcripts have also been detected in human thymocytes, but not in mature T cells, implicating that Blk may play an important role in thymopoiesis (6). Blk function may be redundant, however, as mice that do not express Blk are not impaired with respect to B cell development and immune response (7).
- Dymecki, S.M. et al. (1990) Science 247, 332-336.
- Wasserman, R. et al. (1995) J. Immunol. 155, 644-651.
- Sefton, B.M. et al. (1994) Curr. Opin. Immunol. 6, 372-379.
- Malek, S.N. et al. (1998) Proc. Natl. Acad. Sci. USA 95, 7351-7356.
- Yao, X.R. et al. (1993) Proc. Natl. Acad. Sci. USA 90, 7946-7950.
- Islam, K.B. et al. (1995) J. Immunol. 154, 1265-1272.
- Texido, G. et al. (2000) Mol. Cell Biol. 20, 1227-1233.
Application References
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Companion Products
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
