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Specifications

N-terminal GST-tagged recombinant human B-Raf residues 416-end, containing a V599E mutation. Expressed by baculovirus in Sf21 insect cells.
Catalog #	B2680-02
Specific Activity	~50,000U/mg
	1 unit of B-Ra=1 unit of MAPK2(m) activity which in turn is equivalent to 1nmol phosphate incorporated into 0.33mg/ml myelin basic protein per minute at 30ºC wtih a final ATP concentration of 100uM. Determined by a triple linked assay which involves the activation of MEK1(h) by B-Raf, followed by the subsequent activation of MAPK2(m) by the activated MEK1(h).
Applications	MS Tryptic Fingerprint: Confirmed identity as B-Raf with ~30% amino acid coverage of the translated native sequence.
Recommended Dilutions	Kinase Assay: 17-34pg was used to activate 0.2uM MEK1, unactive. Activated MEK1 was in turn used to activate MAP Kinase 2/Erk2, unactive. Activation of MAP Kinase 2/Erk2 was demonstrated by phosphorylation of MBP substrate in vitro.
	Optimal dilutions to be determined by the researcher.
Storage and Stability	Aliquot to avoid repeated freezing and thawing and store at -70°C. Aliquots are stable for at least 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Molecular Weight	67.3kD
Source	Recombinant protein expressed in Sf21 cells
Purity	Glutathione agarose; ~48% by SDS-PAGE and Coomassie blue staining.
Concentration	~0.1mg/ml
Form	Supplied as a liquid in 50mM Tris-HCl, pH 7.5, 150mM sodium chloride, 0.1mM EGTA, 0.03% Brij-35, 270mM sucrose, 1mM benzamidine, 0.2mM PMSF, 0.1% 2-mercaptoethanol.
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.