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HTRF KRAS WT GTP Binding Kit, 500 Assay Points

The KRAS WT GTP binding kit is designed to identify KRAS WT / GTP inhibitors.

For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption & disposal requirements under European REACH regulations (EC 1907/2006).
Product Variants
Part number: 64BDKRASWPEG
Unit Size: 500 Assay Points
List price: USD 1,254.00
Your price:
USD 1,254.00
USD 1,254.00 /each
Part number: 64BDKRASWPEH
Unit Size: 10,000 Assay Points
List price: USD 15,010.00
Your price:
USD 0.00
USD 15,010.00 /each

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
500
Assay Target Type
Kit
Assay Technology
HTRF
Brand
HTRF
Quantity
1
Shipping Conditions
Shipped in Dry Ice
Therapeutic Area
Oncology & Inflammation
Unit Size
500 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.

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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.

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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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4assay-validation-human-kras-wt-gtp-binding-2.svg

 

Resources

1-2 of 2 Resources
Guide
HTRF solutions, guide to major applications

This guide provides you an overview of HTRF applications in several therapeutic areas.

Whitepaper
KRAS biomarker becomes a target of choice in certain cancer therapies

Get the latest insights on KRAS and see how it has switched from being an undruggable to a druggable cancer target.

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SDS, COAs, Manuals and more Illuminator

SDS, COAs, Manuals and more

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