BRAF (V600E), GST-His-Tag Recombinant

Catalog #

40533

$355 *

Size: 10 µg

Qty

Bulk/Custom

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Description

Recombinant human BRAF (B-Raf proto-oncogene, serine/threonine kinase), encompassing amino acids 417-766(end) containing the mutation of interest (V600E). This construct contains an N-terminal GST-tag followed by a His-tag (6His). A Thrombin Cleavage Site to facilitate removal of the GST portion is also present between the His portion and the protein. The recombinant protein was affinity purified.

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Synonyms

BRAF1, RAFB1, B-raf 1, MGC126806, MGC138284, BRAF (V600E)

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

BRAF Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

BRAF (V600E) (GST-His-417-766(end))

Host Species/Expression System

Sf9

Purity

≥79%

Format

Aqueous buffer solution

Formulation

40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 0.04% Tween-20, 20% glycerol, and 3 mM DTT

69 kDa

Amino Acids

417-766(end)

Specific Activity

166 pmol/min/μg

Genbank #

NM_004333

UniProt #

P15056

Tag(s)

N-terminal GST-His-tag

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 600 within the activation segment of B-RAF (2).

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.

Storage/Stability

At least 6 months at –80°C

Assay Conditions

Assay was done in Kinase buffer containing 1 mM DTT, 0.1 mg/ml Raf substrate (Mek1 BPS Bioscience #40075 Lot 190508-3), and 10 µM ATP. Reaction was done at 30°C for 45 min. Amount of ATP transferred was calculated using Kinase-Glo reagent (Promega).

Instructions for Use

Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.

Applications

Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Shipping Temperature

-80°C

Warnings

Avoid freeze/thaw cycles

Scientific Category

Kinase/Serine-Threonine

Regulatory Status

Research Use Only

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Datasheets/Protocols

40533 Lot 190501-2

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BRAF (V600E), GST-His-Tag Recombinant

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