The RAF family of signaling proteins has three isoforms: A RAF, RAF 1 and B RAF. These proteins are instrumental in forwarding the stimulus generated by mitogens from monomeric G proteins to the MAP kinase pathway. RAF 1 is known to be downstream of Ras and binds to it by an N terminal noncatalytic domain. This domain is conserved in A RAF and B RAF. 14-3-3 is also known to be an activator of RAFs. Activated RAF directly phosphorylates and activates MEK. A RAF is expressed at a very high concentration in urogenital and kidney tissues.
Recombinant protein corresponding to human A-RAF, aa282-606 (end), with N-terminal GST tag and C-terminal His tag, containing the mutations YY301-302DD, expressed in Baculovirus infected Sf9 cell expression system.
Specific Activity:40pmol/min/ug
Assay Conditions:40mM Tris, 10mM MgCl2, M2865-06J (5ug), 1uM ATP and 4-8ng/ul R0495-05A at 30ºC for 30 minutes. ATP reduction is detected using Kinase-Glo Luminescent Kinase Assay Platform.
Application:Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling. Other applications not tested.
Recommended Dilution:Optimal dilutions to be determined by the researcher.
Storage and Stability:Aliquot to avoid repeated freezing and thawing and store at -70°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Aliquots are stable for at least 6 months.
