Product Pathways - Kinases
Lck Kinase #7487
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 Lck (Met1-Pro509) 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 full-length human Lck (Met1-Pro509) (GenBank Accession No. NM_005356) 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 Lck protein was analyzed using SDS/PAGE followed by Coomassie stain.
Kinase Assay - Radiometric
Figure 2. Lck kinase activity was measured in a radiometric assay using the following reaction conditions: 5 mM MOPS, pH 7.2, 2.5 mM β-glycerophosphate, 1mM EGTA, 0.4 mM EDTA, 5 mM MgCl2, 0.05 mM DTT, 50 μM ATP, Substrate: Poly (EY 4:1) 400 ng/μl, and variable amounts of Lck kinase.
Quality Control
The theoretical molecular weight of the Lck protein is 84 kDa. The purified kinase was quality controlled for purity using SDS-PAGE followed by Coomassie stain [Fig.1]. Lck kinase activity was determined using a radiometric assay [Fig.2].
Background
Lck belongs to the Src-like non-receptor tyrosine kinase family with the typical Src family kinase structure: a unique amino terminal domain (Src homology 4 domain, SH4) followed by an SH3 domain, an SH2 domain, a kinase domain (SH1), and a carboxy-terminal negative regulatory domain (1). Lck activity is controlled by the interactions of SH2 and SH3 domains as well as tyrosine phosphorylation status of the activation loop (2,3). Lck is recruited to the T cell receptor (TCR) complex upon stimulation and activates downstream tyrosine kinases to initiate T cell signaling (4). Lck is also found to be involved in the regulation of mitochondrial apoptosis pathways and may be responsible for some anticancer drug induced apoptosis (5,6).
- Palacios, E.H. and Weiss, A. (2004) Oncogene 23, 7990-8000.
- Mustelin, T. and Tasken, K. (2003) Biochem J 371, 15-27.
- Gervais, F.G. et al. (1993) Mol Cell Biol 13, 7112-21.
- Straus, D.B. and Weiss, A. (1992) Cell 70, 585-93.
- Belka, C. et al. (2003) Oncogene 22, 176-85.
- Gruber, C. et al. (2004) Biochem Pharmacol 67, 1859-72.
Application References
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Companion Products
- 9805 HTScan® Tyrosine Kinase Buffer (4X)
- 9804 ATP (10 mM)
- 7450 Tyrosine Kinase Substrate Screening Kit
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.
