KRAS WT GTP binding kit
KRAS WT/GTP biochemical binding assay
The KRAS / GTP binding Standard is intended for use with the KRAS WT/GTP and KRAS G12C/GTP binding kits.
KRAS GTP standard can be used as a standard to control assay reproducibility. The KRAS GTP Standard is intended for use with the KRAS WT GTP binding kit and the KRAS G12C GTP binding kit.
The HTRF KRAS WT or G12C GTP binding assays are in competitive assay formats, using a GTP-Red reagent as KRAS ligand, a 6His tagged human KRAS WT or G12C protein, and an anti 6His Eu Cryptate-labeled antibody. The compound being tested competes with the GTP Red reagent, and thereby prevents FRET from occurring.
The Human KRAS WT or G12C binding assays can be run in a 96- or 384-well low volume white plate (20 µL final). As described here, samples or standards are dispensed directly into the assay plate. The human His-tagged KRAS WT or G12C protein is then added, followed by the dispensing of the HTRF reagents: The anti 6His antibody labeled with Europium cryptate and the GTP-Red reagent. The reagents labelled with HTRF fluorophores may be pre-mixed and added in a single dispensing step. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally.
The KRAS GTP standard competes with the GTP-Red reagent for KRAS WT and KRAS G12C proteins in a correlated manner .
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