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KRAS/SOS1 binding Standard HTRF®

The KRAS/SOS1 binding Standard is intended for use with the KRAS WT/SOS1 and KRAS G12C/SOS1 binding kits.

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  • Developed and tested to Cisbio's high quality standards Developed and tested to Cisbio's high quality standards

The KRAS/SOS1 binding Standard is intended for use with the KRAS WT/SOS1 and KRAS G12C/SOS1 binding kits.

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Overview

KRAS/SOS1 binding standard can be used as a standard to control assay reproducibility. KRAS/SOS1 binding standard is intended for use with the KRAS WT/SOS1 and the KRAS G12C/SOS1 binding kits.

Benefits

  • PRODUCED BY MANUFACTURER
  • STRINGENT QUALITY CONTROL

Assay principle

The KRAS/SOS1 PPI assays include tagged human recombinant partners  (KRAS WT or G12C and SOS1) and labelled anti-tag reagents for HTRF detection. Without any inhibitor, KRAS 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/SOS1 inhibitors or GTP competitors, the HTRF signal decreases.

Principle of the HTRF KRAS G12C/SOS1 binding assay

Assay protocol

The Human KRAS/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 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 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 protocol of the HTRF KRAS WT/SOS1 binding assay

Standard curves with human KRAS WT and G12C

The KRAS/SOS1 binding standard inhibits the KRAS/SOS1 interaction for KRAS WT and KRAS G12C proteins in a correlated manner.

Validation of the HTRF KRAS/SOS1 standard with hKRAS WT and hKRAS G12C

Best practices for pharmacological characterization of PPI inhibitors

Easy pharmacological characterization of PPI modulators. - Technical Notes

HTRF assays handle low- to high affinity protein-protein interactions

Deciphering low- and high affinity interactions - Application Notes

Nuclear receptor ligand identification with HTRF

Monitoring nuclear receptor binding with HTRF assays - Application Notes

HTRF addresses large protein-protein interaction complexes

Challenge large complexes with HTRF assays - Application Notes

HTRF PPI your dream assay served on a plate

Sandwiches aren't just for eating - Infographics

A brief history of Protein-Protein Interactions

How well do you know PPI? - Infographics

Virology research solutions using HTRF Protein-Protein Interaction assays

See how peer researchers challenge the viral life cycle with PPI assays - Application Notes

Plate Reader Requirement

Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.

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