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HTRF KRAS WT / GTP Binding Kit, 10,000 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 and disposal requirements under European REACH regulations (EC 1907/2006).

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Feature	Specification
Application	Protein-Protein Interaction
Sample Volume	2 µL

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

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Product Variants

Unit Size: 500 Assay Points

Part #:

64BDKRASWPEG

List Price

USD 1,254.00

Unit Size: 10,000 Assay Points

Part #:

64BDKRASWPEH

List Price

USD 15,010.00

Product information

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

Other Specifications

Application	Protein-Protein Interaction
Brand	HTRF
Detection Modality	HTRF
Product Group	Kit
Sample Volume	2 µL
Shipping Conditions	Shipped in Dry Ice
Target Class	Binding Assay
Technology	TR-FRET
Therapeutic Area	Oncology & Inflammation
Unit Size	10,000 Assay Points

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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

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.