Kras (G13D)/cRAF/CYPA/Inhibitor Assay Kit

The Kras (G13D) Inhibitor assay kit is a TR-FRET based assay, which is designed to screen Kras inhibitors and determine the Kras-inhibitor binding affinity. Tag2-Kras (G13D) in this assay kit is loaded with GppNHp, which represents the activated Kras. The Ras binding domain (RBD) of cRAF in the kit has a Tag1 at N-terminus. A Terbium-labeled anti-Tag2 antibody binding to the Tag2-Kras serves as a fluorescence donor (HTRF donor), activation of which results in fluorescence resonance energy transfer (FRET) if Tag1-cRAF binds to the Kras, since the binding brings Terbium on the anti-Tag2 antibody close to the fluorophore on the anti-Tag1 antibody (HTRF acceptor). Thus, the binding status can be quantitively measured by calculating the ratio of the emission fluorescence intensity of the acceptor (665 nm) and donor (620 nm). If an inhibitor associated with CYPA binds to the Kras and blocks the cRAF binding, the HTRF signal will be reduced.

Please contact us at for specific academic pricing.

Background

Kras is a member of the RAS protein family, which are a class of small GTPases involved in cell signaling pathways. The Ras signaling pathway regulates diverse cellular processes, including cell proliferation, differentiation, and survival. Conversion of Ras from the inactive GDP-bound state to the active GTP-bound state activates the downstream effector and promotes cell growth. RAF is a key downstream effector of RAS. Since the frequently mutated Ras genes are associated with various human tumors, the Ras-RAF signaling pathway is considered an important therapeutic target for cancer treatment. However, Ras is considered undruggable since it lacks suitable binding pockets on the surface. Recently, a discovery of a small molecule inhibitor blocks Ras-RAF signaling pathway by remolding Cyclophilin A (CYPA) and forming a CYPA:drug:KRAS ternary complex. This inhibitory strategy provides a new method for developing drugs targeting Kras for treatment of cancers.