B-Raf Active Protein
B-Raf (V599E), active Recombinant
Purity
PurifiedGlutathione agarose; ~48% by SDS-PAGE and Coomassie blue staining.
Synonyms
B-Raf; B-Raf (V599E); active Recombinant; B-Raf active protein
Purity/Purification
Purified
Glutathione agarose; ~48% by SDS-PAGE and Coomassie blue staining.
Glutathione agarose; ~48% by SDS-PAGE and Coomassie blue staining.
Form/Format
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.
Application Notes
MS Tryptic Fingerprint: Confirmed identity as B-Raf with ~30% amino acid coverage of the translated native sequence.
Dilution: 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.
Dilution: 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.
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).
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).
Preparation and Storage
Aliquot to avoid repeated freezing and thawing and store at -70 degree 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.
Related Product Information for B-Raf active protein
N-terminal GST-tagged recombinant human B-Raf residues 416-end, containing a V599E mutation. Expressed by baculovirus in Sf21 insect cells.
Product Categories/Family for B-Raf active protein
NCBI and Uniprot Product Information
Similar Products
Product Notes
The B-Raf (Catalog #AAA637034) is an Active Protein and is intended for research purposes only. The product is available for immediate purchase. MS Tryptic Fingerprint: Confirmed identity as B-Raf with ~30% amino acid coverage of the translated native sequence. Dilution: 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. Researchers should empirically determine the suitability of the B-Raf for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "B-Raf, Active Protein" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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