KRAS WT/SOS1 binding kit
KRAS WT/SOS1 biochemical binding assay
The KRAS/SOS1 binding Standard is intended for use with the KRAS WT/SOS1 and KRAS G12C/SOS1 binding kits.
KRAS/SOS1 binding standard can be used as a standard to control assay reproducibility. KRAS/SOS1 binding standard is intended for use with the KRAS WT/SOS1 and the KRAS G12C/SOS1 binding kits.
The KRAS/SOS1 PPI assays include tagged human recombinant partners (KRAS WT or G12C and SOS1) and labelled anti-tag reagents for HTRF detection. Without any inhibitor, KRAS loaded with GTP binds to SOS1 , and the binding of each detection reagent to its tagged target generates an HTRF signal. In the presence of KRAS/SOS1 inhibitors or GTP competitors, the HTRF signal decreases.
The Human KRAS/SOS1 binding assay can be run in a 96- or 384-well low volume white plate (20 µL final). As described here, compounds or standards are dispensed directly into the assay plate. The pre-mixed GTP and human Tag1-KRAS protein is then added, together with the human Tag2-SOS1 protein, followed by the dispensing of the HTRF reagents, the anti-Tag2 antibody labeled with Terbium cryptate and the anti-Tag1 antibody labeled with XL665. 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/SOS1 binding standard inhibits the KRAS/SOS1 interaction for KRAS WT and KRAS G12C proteins in a correlated manner.
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