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HTRF KRAS WT GTP BIND. KIT - 500 PTS


品牌: Revvity









产品介绍
产品信息
Overview
A fast and easy way to identify new KRAS WT / GTP competitors.KRAS is a small GTPase implicated in various biological processes, such as cell proliferation, cell survival, and cell metabolism. This proto-oncogene is well known to be mutated in many cancer subtypes, inducing an uncontrolled proliferation and cell metabolism modifications. It thereby contributes to the Warburg effect in cancer cells. Like the majority of small GTPases, KRAS binds to GDP in its inactive form or binds to GTP to switch to the active form. KRAS G12C is one of the most commonly present mutant forms in cancer which lead to a permanently active state of KRAS. Identifying new KRAS / GTP competitors is therefore a relevant strategy to control biological processes involved in cancer growth by reducing the KRAS activity, as well as the associated pathways.
Specifications
Assay Points | 10000 |
---|---|
Assay Target Type | Kit |
Assay Technology | HTRF |
Brand | HTRF |
Quantity | 1 |
Shipping Conditions | Shipped in Dry Ice |
Therapeutic Area | Oncology & Inflammation |
Unit Size | 10,000 Assay Points |
Video gallery


How it works
Assay principle
The HTRF KRAS GTP binding assay is in a competitive assay format, using a GTP-Red reagent as KRAS WT ligand, a 6His tagged human KRAS WT protein, and an anti 6His Cryptate-labeled antibody. The compound being tested competes with the GTP Red reagent, and thereby prevents FRET from occurring.
Assay protocol
The Human KRAS WT binding assay 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 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 labeled 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.
Assay details
GTP-Red reagent Kd | 70nM
|
---|---|
GTP-Red reagent concentration | 50nM
|
KRAS GTP binding Standard (GDP) IC50 | 30nM
|
KRAS GTP binding Standard Ki
| 20.4nM
|
Assay validation
Screening of GTP competitors with human KRAS WT protein
Various nucleotides known to be GTP competitors were added to the assay. GDP (assay standard), GTPyS, and GTP displayed the right potency, in good correlation with the literature. ATP, a non-specific nucleotide, and BAY-293, a KRAS/SOS1 inhibitor, had no effect on the assay, confirming the specificity of the kit.
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