Sandwich ELISA performed on the lysates of 293T cells overexpressing BRAF V600E or BRAF wild type His-tag proteins using BRAF V600E antibody (clone RM8), shows the specificity of RM8 to the V600E mutation. His-tag fusion proteins were captured by an Anti-His-Tag antibody.
BRAF belongs to the raf/mil family of serine/threonine protein kinases. It plays a role in regulating the MAP kinase/ERKs signaling pathway, which affects cell division, differentiation, and secretion (Entrez Gene).
The BRAF V600E mutation results in an amino acid substitution at position 600 in BRAF, from a valine (V) to a glutamic acid (E). This mutation occurs within the activation segment of the kinase domain. Approximately 80–90% of V600 BRAF mutations are V600E (COSMIC; Lovly et al. 2012; Rubinstein et al. 2010). Mutations at V600 result in increased kinase activity and are transforming in vitro. BRAF mutations are usually found in tumors with no driver mutations in NRAS, KIT, and other genes (MyCancerGenome.org).
|Immunogen:||A peptide corresponding to BRAF V600E mutant|
|Synonyms:||Serine/threonine-protein kinase B-raf, Proto-oncogene B-Raf, BRAF1, RAFB1, p94|
|Observed MW:||Refer to figure|
|Specificity:||This antibody reacts to the BRAF V600E mutant. No cross reactivity with wild type BRAF.|
|Applications:||IHC, ICC, ELISA, WB|
|Recommended dilutions:||IHC, ICC: 0.5 – 5 ug/mL
ELISA: 0.5 – 2 ug/mL
WB: 0.5 – 2 ug/mL
|Purification Method:||Protein A affinity purified from an animal origin–free culture supernatant|
|Buffer:||50% Glycerol/PBS with 1% BSA and 0.09% sodium azide|
|Storage Conditions:||Stable for 1 year at -20.0°C from date of receipt.|