50 mM Tris-HCl pH 7.5, 10 mM DTT, 0.1 mM EGTA, 10 mM MgAc
Three step assay in which B-Raf activates unactive MKK1, which in turn activates unactive p42MAPKinase. Activity of p42MAPkinase is then assayed against myelin basic protein as substrate (final conc. of 0.3 mg/ml), in a standard filter binding assay.
Calculated molecular mass:
Mono-Isotopic Mass: 111,727.51
Average Mass: 111,798.57
50 mM Tris-HCl pH 7.5, 270 mM Sucrose, 150 mM NaCl, 0.1 mM EGTA, 0.1 % 2-mercaptoethanol, 0.03 % Brij-35, 0.2 mM PMSF, 1 mM Benzamidine
Gel Information :
Gel Image 1:
Amino acids A2 – H766 (end) of human B-Raf. Residue A238 of the fusion protein is equivalent to A2 of the native enzyme. The GST tag is located at residues 1 - 220.
PreScission (LEVLFQGP) residues 221 - 228
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