N-terminal GST-tagged recombinant human B-Raf residues 416-end, containing a V599E mutation. Expressed by baculovirus in Sf21 insect cells.
Specific Activity: ~50,000U/mg1 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.
仕様
Size：5ug
Source Antigen：Recombinant protein expressed in Sf21 cells
Grade：Purified
Purity：Glutathione agarose; ~48% by SDS-PAGE and Coomassie blue staining.
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.
