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

PubChem ID

137278711

Name

AMG510

Measurement

>100µM - IC50

PubChem ID

137322663

Name

BAY-293

Measurement

1µM - IC50

PubChem ID

138911318

Name

BI-3406

Measurement

0.58µM - IC50

PubChem ID

134326084

Name

MRTX1257

Measurement

3.8µM - IC50

PubChem ID

138756246

Name

BI-2852

Measurement

4.4µM - IC50

PubChem ID

168268166

Name

BI-2865

Measurement

0.013µM - IC50

Clinical Relevance

Therapeutic Area:

Oncology/Immuno-Oncology

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei

RELATED SOLUTIONS

Item: PP320