K-Ras Binding Assay Service (Thermal shift)

KRAS-related Assays: K-Ras binding assay service for compound screening and profiling using the thermal shift assay format to measure compound binding to the target

Target

K-Ras

HGNC

KRAS

Protein

KRAS aa 2-169, N-terminal his tag. Expressed in E. coli.

Synonyms

KRAS, C-K-RAS, CFC2, K-RAS2A, K-RAS2B, K-RAS4A, K-RAS4B, KI-RAS, KRAS1, KRAS2, NS, NS3, RASK2

Scientific information

KRas: P01116 (https://www.uniprot.org/uniprot/P01116)

Screening location

Malvern, PA, US

Reaction

Compound binding to K-Ras

Readout

Protein stability change due to ligand binding via thermal shift assay

Concentration of protein in assay

The typical protein concentration in assay is 0.15 mg/ml

Concentration of compound in assay

Variable, 25 µM is recommended

Apo form Tm (°C)

50.1

Reference Compound ΔTms
Construct ARS-1620 AMG-510 BI-2852 BAY-293
Wild type 0 0.5 0 -0.5
G12C peak 1 4.7 3.3 0.5 0.25
G12C peak 2 12.5 16.5 no 2nd peak no 2nd peak
G12D 0.5 0.5 1 0.5
G12V 0.25 0.25 0.75 0

 

Note: 0.5°C is a typical noise observed in thermal shift assay which is not significant.

Reference compound testing in thermal shift assay with K-RAS wild type and mutants

KRAS_thermalshift_RAS_Malvern_WT
KRAS_thermalshift_RAS_Malvern_G12C
KRAS_thermalshift_RAS_Malvern_G12D
KRAS_thermalshift_RAS_Malvern_G12V