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Encompassing Amino Acids: 417-end
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: Assay was performed in a kinase buffer containing 0.1 mg/ml inactive MEK1 substrate (BPS Cat# 40075) and 10 µM ATP. Reaction was done at 30°C for 30 minutes. ATP transferred was calculated using Kinase-Glo® (Promega).
Background: BRAF is a member of the RAF family that is activated by members of the Ras family upon growth factor-induced stimulation. Active Ras can induce heterodimerization of cRaf and BRAF and this may explain the observed cooperativity of cRaf and BRaf in cells responding to growth factor signals (1). Activating mutations in the BRAF gene are present in a large percentage of human malignant melanomas and in a proportion of colon cancers. The vast majority of these mutations result in a valine to glutamic acid change at residue 599 within the activation segment of B-RAF (2).
Description: Human v-raf murine sarcoma viral oncogene homolog B1 (BRAF), GenBank Accession No. NM_004333, a.a. 417-end with Val-to-Glu mutation on a.a. 600* and N-terminal GST-tag and His-tag, MW=69 kDa, expressed in Sf9 insect cells via a baculovirus expression system.*This mutation is often cited as V600E.
Format: Aqueous buffer solution
Formulation: 40 mM Tris, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 3 mM DTT and 20% glycerol.
Genbank: NM_004333
Host Cell Line: Sf9 cells
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag, N-terminal His-tag
Uniprot: P15056
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Weber, C.K., et al., Cancer Res. 2001 May 1, 61(9): 3595-8.
2. Mercer, K.E., et al., Biochim Biophys Acta. 2003 Jun 5, 653(1): 25-40.
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