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HTRF KRAS G12C/SOS1 binding kit, 500 Assay Points

The KRAS G12C/SOS1 binding kit is designed to identify KRAS G12C/SOS1 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: 64KRASG12PEG
Unit Size: 500 Assay Points
List price: USD 1,254.00
Your price:
USD 1,254.00
USD 1,254.00 /each
Part number: 64KRASG12PEH
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 inhibitors of KRAS G12C/SOS1 protein interactions.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. The Ras guanine nucleotide exchange factor, also called SOS1, is a GEF protein promoting the active form of KRAS. The upregulation of the KRAS/SOS1 ction leads to cancer phenotypes. ying new KRAS/SOS1 inhibitors or GTP competitors refore the two major ies rol 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 KRAS G12C / SOS1 PPI assay includes tagged human recombinant partners (KRAS G12C and SOS1) and labeled anti-tag reagents for HTRF detection. Without any inhibitor, KRAS G12C 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 G12C / SOS1 inhibitors or GTP competitors, the HTRF signal decreases.

PPI how it works assay principle KRAS G12C sos1

 

 

Assay protocol

The Human KRAS G12C / 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 G12C 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 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.

PPI how it works assay protocol KRAS G12C sos1

 

Assay validation

KRAS G12C/SOS1 inhibitor screening

Various compounds known to be KRAS or SOS1 inhibitors were added to the assay. BAY-293, BI-3406, BI-2852, MRTX-1257, and the two KRAS G12C selective compounds, ARS-1620 and AMG-510, displayed the right potency and pharmacological ranking in good correlation with the literature. ARS-1620 and AMG-510 results confirm that the KRAS G12C/SOS1 binding kit is specific to G12C mutated form of KRAS.

Assay validation KRAS G12C SOS1 3

 

Competitive or non competitive nucleotide screening

The two nucleotides GDP and ATP were added to the assay. GDP, a GTP competitor well known to bind KRAS G12C, displayed the right potency in good correlation with the literature. Meanwhile ATP, a non GTP competitive nucleotide, had no effect on the assay, thus confirming the specificity of the kit in detecting only GTP competitors or KRAS G12C/SOS1 inhibitors.

Assay validation KRAS G12C SOS1 2

 

Resources

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Guide
HTRF solutions, guide to major applications

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

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SDS, COAs, Manuals and more

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