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Encompassing Amino Acids: 30-397
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: 100 µl of 240 nM biotinylated MEK1 peptide was coated onto a neutravidin plate for 1 hour at 37°C. After overnight blocking with TBS superblock, the reaction was performed with 10 µM ATP for 60 minutes at RT, and phospho-MEK1 was detected using an anti-phospho-MEK1 antibody. Relative specific activity was determined using a standard kinase under the same conditions.
Background: COT is an oncogene that can activate both the MAP kinase and JNK kinase pathways. COT activates IkappaB kinases and induces the nuclear production of NF-κB. C-terminal catalytic domain of KSR2 associates with COT and KSR2 can negatively regulates the kinase activity of COT in vitro. Co-transfection of KSR2 with COT in cells lead to reduced COT-mediated ERK activation and COT-induced IL8 production in a dose-dependent manner. Cot is one of the MAP kinase kinase kinases that regulates the ERK1/ERK2 pathway in response to IL-1. Blockage of expression of Cot results in failure of IL-1 to induce an increase in IL-8 and MIP-1betamRNA levels.
Description: Human Cot/Tpl2 (GenBank Accession No. NM_005204), (a.a. 30-397), with N-terminal GST-His-tag, MW= 72 kDa, expressed in a Baculovirus infected Sf9 cell expression system.
Format: Aqueous buffer solution
Formulation: 25 mM Tris-HCl, pH 8.0, 100 mM NaCl, 0.05% Tween-20, 50% glycerol, and 3 mM DTT.
Genbank: NM_005204
Host Cell Line: Sf9 cells
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-His-tags
Uniprot: P41279
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Stafford, M.J. et al., FEBS Lett. 580, 4010-4014 (2006).
2. Babu, G. et al., J. Biol. Chem. 281, 14041-14047 (2006).