KRAS Screening & Profiling Services
KRAS is an important protein in the RAS/MAPK signaling pathway that transmits signals from activated cell surface receptors leading to cell proliferation and survival. KRAS function is controlled by cycling through an inactive, GDP-bound state and an active, GTP-bound state. Activation of KRAS occurs when it binds to Guanine Nucleotide Exchange Factor (GEF) proteins, including SOS, which initiate the exchange of GDP for GTP. Active, GTP-bound KRAS can trigger activation of downstream signaling molecules including RAF, PI3K, and RAL. KRAS can self-inactivate by cleaving the terminal phosphate group on the GTP, converting it to GDP. This inactivation process is accelerated by GTPase-activating (GAP) proteins.
KRAS has emerged as a target of significant clinical interest, due to mutations in the KRAS protein that promote the pathogenesis of 20-30% of human cancers. Testing a patient for the presence of specific KRAS mutants serves as a highly predictive marker for success rates to various types of lung and colon cancer therapies. The development of inhibitors against KRAS G12C, G12D, and G12V holds a promising future for oncology.
![KRAS Screening and Profiling Services](https://bpsbioscience.com/media/wysiwyg/Landing_Pages/KRAS_diagram.png)
We offer wild-type and mutant KRAS screening and profiling services to identify specific inhibitors. Our team of experts, using our in-house developed biochemical assay kits, can provide:
- Screening for activity across your compound library
- Determination of IC50
- Project guidance and questions answered in a timely manner
- Delivery of detailed results promptly and on time
If follow-up studies are required, we can provide the same active proteins as the ones used in our assays.
Learn more about the process and deliverables.
Fluorogenic Nucleotide Exchange Assay
![Fluorogenic Nucleotide Exchange Assay](https://bpsbioscience.com/media/wysiwyg/Landing_Pages/nucleotide_exchange_diagram.png)
Fluorogenic Nucleotide Exchange Assays are homogeneous assays designed for the screening and profiling of KRAS antagonists/inhibitors. In this assay, KRAS is pre-loaded with fluorescent BODIPY-GDP. Adding GTP in the presence of EDTA displaces BODIPY-GDP. The fluorescence intensity decreases as the BODIPY-GDP is displaced by GTP. Several KRAS inhibitors lock KRAS in the (inactive) GDP-bound conformation and prevent GDP/GTP exchange. In this scenario, the fluorescence intensity increases with drug concentration as more BODIPY-GDP remains bound to KRAS.
AlphaLISA® Coupled Nucleotide Exchange Assay
![AlphaLISA® Coupled Nucleotide Exchange Assay](https://bpsbioscience.com/media/wysiwyg/Landing_Pages/Coupled_exchange_diagram.png)
AlphaLISA® Coupled Nucleotide Exchange Assays are designed for the screening and profiling of KRAS antagonists/inhibitors by monitoring the binding of an effector protein such as the Ras binding domain of Raf1, (RBD-cRaf) to active KRAS (G12C, in this example). First, a sample containing GDP-loaded KRAS(G12C) is incubated with SOS1 and GTP for the nucleotide exchange. Next, RBD-cRAF is added and incubated for the effector-RAS binding. Then, acceptor and donor beads are added and incubated for detection followed by reading the Alpha-counts.
Available Service Assays
Note: Not all targets will have a commercially available reference control. Please inquire with our team if you need to furnish your own controls for specific targets.
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│Related Services
- Cell Signaling Pathway Screening & Profiling
- Kinase Screening & Profiling
- Custom Biochemical Assay Development