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Recombinant Protein - c-Raf [Y340D, Y341D] (306 - 648) *

Name: c-Raf [Y340D, Y341D] (306 - 648) *
Price per aliquot (100µg): £100.00
Catalog Number (DU Number):
DU811
Accession:
BC018119
Expression:
baculovirus
Tag and Terminus:
N-Term
GST
Uncleaved
Purification Method:
GSH Sepharose
Enzymatic Assay Format:
hree step assay in which c-Raf activates unactive MKK1 [DU 1843], which in turn activates unactive p42MAPKinase [DU 650 or DU 1844]., Activity of p42MAPkinase is then assayed against MBP as substrate (final concentration of 0.3 mg/ml), in the standard filter binding assay.
Enzymatic Buffer:
50 mM Tris-HCl pH 7.5, 0.1 mM EGTA, 0.1 % 2-mercaptoethanol, 0.2 mM sodium vanadate, 0.5 μM microcystin-LR, 10 mM magnesium acetate
Calculated molecular mass:
Mono-Isotopic Mass: 65, 727.62 daltons
Average Mass: 65, 769.87 daltons
Protein Activity:
Constitutively Active
Purity:
>80 %
Storage Buffer:
50 mM Tris-HCl pH 7.5, 270 mM Sucrose, 150 mM NaCl, 0.1mM EGTA, 0.1 % 2-mercaptoethanol, 0.02 % Brij-35
Storage Temperature:
-70 °C
Theoretical pI:
8.2
Gel Information :
  • Gel Image 1: 
Native Sequence:
Amino acids S306 – F648 (end) of human c-Raf. Residue S232 of the fusion protein is equivalent to S306 of the native enzyme. The enzyme has a Y430D and a Y341D mutation in order to mimic phosphorylation of the enzyme. Residues Y430 and Y431 are equilivalent to D266 and D267 of the fusion protein. The GST tag is located at residues 1 – 220.
Protease Cleavage:
PreScission (LEVLFQGPL) at residues 221 – 229
Cloning Sites:
BamHI and EcoRI site of pFastBAC GST

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