Revvity, 64KRASTPEG, HTRF Human Total KRAS Detection Kit, 500 Assay Points

Overview
KRAS is a monomeric GTPases that relays signals from receptor tyrosine kinases promoting cell proliferation, differentiation or survival. KRAS is one of the most frequently mutated oncogenes in cancer. Mutation is commonly associated with poor prognosis and resistance to therapy. The KRAS G12C mutation is one of the more common KRAS mutations. KRAS G12C is constitutively active, in a persistently GTP-bound state, resulting in overstimulation of the KRAS effector signaling MAPK pathways and driving tumor growth.
How it works
Total KRAS assay principle
The Total KRAS assay quantifies the expression level of KRAS in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total KRAS assay uses two labeled antibodies, one coupled to a donor fluorophore and the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In the presence of KRAS in a cell extract, the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the protein present in the sample, and provides a means of assessing the protein's expression under a no-wash assay format.

 

Total KRAS two-plate assay protocol
The two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates into a 384-well low volume detection plate before the addition of HTRF Total KRAS detection reagents. This protocol allows for the cells' viability and confluence to be monitored. 

Total KRAS one-plate assay protocol
Detection of Total KRAS with HTRF reagents can be performed in a single plate used for culturing, stimulation, and lysis. No washing steps are required. This HTS designed protocol facilitates miniaturization while maintaining robust HTRF quality. Application-Cell Signaling
Brand-HTRF
Buffer/Solvent-Lysis Buffer 6
Detection Modality-HTRF
Host Species-Human
Lysis Buffer Compatibility-Lysis Buffer 1 Lysis Buffer 4
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target-KRAS
Target Class-Phosphoproteins
Target Species-Human
Technology-TR-FRET
Therapeutic Area-Inflammation Oncology
Unit Size-500 assay points