HTRF KRAS G12C GTP Binding Kit, 500 Assay Points

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HTRF KRAS G12C GTP Binding Kit, 500 Assay Points

The KRAS G12C GTP binding kit is designed to identify KRAS G12C / 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 information

SDS, COAs, Manuals and more

Product Variants

partnumber

Part number: 64BDKRASGPEG

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Unit Size: 500 Assay Points

List price: USD 1,254.00

Your price:

USD 1,254.00

USD 1,254.00 /each

partnumber

Part number: 64BDKRASGPEH

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Unit Size: 10,000 Assay Points

List price: USD 15,010.00

Your price:

USD 0.00

USD 15,010.00 /each

Product information

Overview

A fast and easy way to identify new KRAS G12C/GTP competitors.KRAS is a small GTPase implicated ous 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 into active form. KRAS G12C is one of the most commonly present mutant forms in cancer which leads to a permanently active state of KRAS. ying new KRAS/ GTP competitors efore a relevant strategy rol biological processes involved in cancer growth, by reducing the KRAS activity as well as the associated pathways.

Specifications

Other 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 which uses a GTP-Red reagent as KRAS G12C ligand, a 6His tagged human KRAS G12C 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 G12C 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 G12C protein is then adde,d 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.

Assay details

GTP-Red reagent Kd	70nM
GTP-Red reagent concentration	50nM
KRAS GTP binding Standard (GDP) IC50	34nM
KRAS GTP binding Standard Ki	19.8nM

GTP-Red reagent Kd

70nM

GTP-Red reagent concentration

50nM

KRAS GTP binding Standard (GDP) IC50

34nM

KRAS GTP binding Standard Ki

19.8nM

Assay validation

Screening of GTP competitors with human KRAS G12C 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.