Biological Information
Background Information:
KRAS encodes a GTPase, K-Ras, acting as a switch, turned on and off by the GTP and GDP. Key effectors downstream of KRAS include MAPK, PI3K, and RalGEF; all of these effectors are responsible for cell cycle regulation, metabolic changes, cell proliferation, survival, and differentiation. Driver mutations in KRAS underlie the pathogenesis of up to 20% of human cancers. One frequent driver mutation, KRASG12C, the most common subtype in non-small cell lung cancer (NSCLC), is adjacent to a shallow binding site. K-Ras inhibitors irreversibly bind with a nucleophilic sulfur atom of Cys-12 and hence selectively target mutant K-Ras and leave wild-type untouched.
Family:
Other
Class:
Other
Protein Name:
KRAS
Uniprot Number:
P01116
Protein Aliases:
K-Ras 2
Gene Name:
KRAS
Gene ID:
3845
Gene Aliases:
KRAS1
Accession Number:
NM_033360
Organism:
Human
Construct Details:
Fragment, Recombinant
Assay Information
Assay Type:
Functional
Functional Mode:
Antagonist
Detection Method:
Spectrofluorimetry
Platform:
Plate reader
Measured Response:
Fluorescence
Testing Information
Procedure Summary:
KRAS WT expressed in E coli cells is used. Test compound and/or vehicle, pretreated KRAS WT/BODIPY-GDP/Terbium-streptavidin (Tb-SA) are combined. Preincubate for 60 minutes at 25°C. Pre-mixed SOS1/GTP are added into each well and incubated for 30 minutes at 25°C. Read plates on a spectrofluorometer (Excitation: 337 nm; Emission 515/486 nm). Compounds are screened at 10 μM.
Substrate:
BODIPY-GDP
Incubation:
30 mins at 25°C
Control Inhibitor:
BI-2865
Criteria For Significance:
≥ 50% of max stimulation or inhibition
Test Concentration / Dose:
Suggested concentration for initial testing is 10 uM
Test Sample Requirements:
Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).
Minimum Order Quantity:
2
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Oncology/Immuno-Oncology
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
RELATED SOLUTIONS