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AURORA BIOLABS LLC Kras G12D CYPA Assay, 384s rxn
Supplier: AURORA BIOLABS LLC 57274123CK384
The Kras (G12D) 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 (G12D) in this assay kit is loaded with GppNHp, which represents the activated Kras. The Ras binding domain (RBD) of cRAF 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.
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