Recombinant Human BRAF fused with MYC/DDK tag at C-terminal was expressed in HEK293.
This gene encodes a protein belonging to the raf/mil family of serine/threonine protein kinases. This protein plays a role in regulating the MAP kinase/ERKs signaling pathway, which affects cell division, differentiation, and secretion. Mutations in this gene are associated with cardiofaciocutaneous syndrome, a disease characterized by heart defects, mental retardation and a distinctive facial appearance. Mutations in this gene have also been associated with various cancers, including non-Hodgkin lymphoma, colorectal cancer, malignant melanoma, thyroid carcinoma, non-small cell lung carcinoma, and adenocarcinoma of lung. A pseudogene, which is located on chromosome X, has been identified for this gene.
25 mM Tris.HCl, pH 7.3, 100 mM glycine, 10% glycerol.
BRAF kinase activity was measured in an HTRF® assay. Varying concentrations of BRAF were added to a reaction mix containing ATP and a biotinylated kinase substrate (HTRF substrate 2) and was incubated at 37°C for phosphorylation. HTRF detection reagents were then added, the reaction was incubated for 30 minutes at room temperature. Time-resolved fluorescent signal (Delta R) was measured on a Flexstation 3 microplate reader.
> 80% as determined by SDS-PAGE and Coomassie blue staining
>50 ug/mL as determined by microplate BCA method