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KRAS / GTP binding Standard HTRF®

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

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Overview

KRAS GTP standard can be used as a standard to control assay reproducibility. The KRAS GTP Standard is intended for use with the KRAS WT GTP binding kit and the KRAS G12C GTP binding kit.

Benefits

  • PRODUCED BY MANUFACTURER
  • STRINGENT QUALITY CONTROL

Assay principle

The HTRF KRAS WT or G12C GTP binding assays are in competitive assay formats, using a GTP-Red reagent as KRAS ligand, a 6His tagged human KRAS WT or G12C protein, and an anti 6His Eu Cryptate-labeled antibody. The compound being tested competes with the GTP Red reagent, and thereby prevents FRET from occurring.

Principle of the HTRF KRAS GTP competitive assay

Assay protocol

The Human KRAS WT or G12C binding assays 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 or G12C 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 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 competitive assay

Validation of the KRAS GTP binding standard

The KRAS GTP standard competes with the GTP-Red reagent for KRAS WT and KRAS G12C proteins in a correlated manner .

Validation of the HTRF KRAS GTP standard with hKRAS WT & hKRAS G12C

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Plate Reader Requirement

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