Product Pathways - Tyrosine Kinase/ Adaptors
Etk/BMX Kinase #7446
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Description
Purified recombinant full length human Etk/BMX (Met1-His675) 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 Etk/BMX (Met1-His675) (GenBank Accession No. NM_001721) 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-Etk/BMX fusion protein was analyzed using SDS/PAGE followed by Coomassie stain.
Kinase Assay - Radiometric
Figure 2. Etk/BMX kinase activity was measured in a radiometric assay using the following reaction conditions: 4 mM MOPS, pH 7.2, 2.5 mM β-glycerophosphate, 10 mM MnCl2, 1 mM EGTA, 0.4 mM EDTA, 4 mM MgCl2, 0.05 mM DTT, 40 ng/μL BSA, 50 μM ATP, Substrate: Poly(Glu-Tyr), 400 ng/μL and recombinant Etk/BMX: variable.
Quality Control
The theoretical molecular weight of the GST-Etk/BMX fusion protein is 104 kDa. The purified kinase was quality controlled for purity using SDS-PAGE followed by Coomassie stain [Fig.1]. Etk/BMX kinase activity was determined using a radiometric assay [Fig.2].
Background
Etk, also known as BMX, is a member of the Bruton's tyrosine kinase (Btk) family (1). It is expressed in a variety of hematopoietic, epithelial and endothelial cells. Etk, like other Btk family members, contains a pleckstrin homology (PH) domain and Src homology SH3 and SH2 domains. It participates in multiple signal transduction pathways (2). Phosphorylation of Tyr566 by Src kinase is required for activation of Etk in vivo (3). In endothelial and epithelial cells, Etk is regulated by FAK through phosphorylation at Tyr40 (4).
- Tamagnone, L. et al. (1994) Oncogene 9, 3683-3688.
- Qiu, Y. and Kung, H.J. (2000) Oncogene 19, 5651-5661.
- Tsai, Y.T. et al. (2000) Mol. Cell. Biol. 20, 2043-2054.
- Chen, R. et al. (2001) Nat. Cell Biol. 3, 439-444.
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.
