Catalog number: 421 - 79862
Product Category: Business & Industrial > Science & Laboratory
Size: 2 x 5 ml
79859
The KRAS(G12C) Nucleotide Exchange Assay Kit is designed for screening and profiling of KRAS(G12C) antagonists/inhibitors by using BODIPY-GDP to monitor the GDP or GTP binding status. The kit can be used with two different protocols for greater flexibility, either titrating the inhibitor with fixed GTP concentration or titrating the GTP with fixed inhibitor concentration._x000D_
78355
The KRAS(G12D) Nucleotide Exchange Assay is a homogeneous assay designed for the screening and profiling of KRAS(G12D) antagonists/inhibitors by using BODIPY-GDP to monitor the GDP or GTP binding status. The kit can be used with two different protocols for greater flexibility, either titrating the inhibitor at a fixed GTP concentration or titrating the GTP at a fixed inhibitor concentration.BODIPY® FL GDP, is a mixed isomer in which the BODIPY® FL fluorophore has been attached to the 2' or 3' position of the ribose ring via a linker. BODIPY® FL dye is a green-fluorescent dye with similar excitation and emission to fluorescein or Alexa Fluor™ 488. It is characterized by a high extinction coefficient and high quantum yield and is relatively insensitive to pH changes. The dye has an excited-state lifetime of 5 nanoseconds or longer, ideal for fluorescence polarization-based assays.
78519
The KRAS(G12V) Nucleotide Exchange Assay is a homogeneous assay designed for the screening and profiling of KRAS(G12V) antagonists/inhibitors by using BODIPY®-GDP to monitor the GDP or GTP binding status. The kit can be used with two different protocols for greater flexibility, either titrating the inhibitor at a fixed GTP concentration or titrating the GTP at a fixed inhibitor concentration.BODIPY® FL-GDP is a mixed isomer in which the fluorophore has been attached to the 2' or 3' position of the ribose ring via a linker. It is a green-fluorescent dye with similar excitation and emission to fluorescein or Alexa Fluor™ 488, characterized by a high extinction coefficient and high quantum yield and is relatively insensitive to pH changes. The dye has an excited-state lifetime of 5 nanoseconds or longer.Assay Principle Illustration: KRAS is activated upon binding GTP, when it undergoes a conformational change. KRAS then returns to a GDP-bound inactive state following the hydrolysis of GTP to GDP. In this assay, KRAS is pre-loaded with fluorescent BODIPY®-GDP and therefore is inactive. Adding GTP in the presence of EDTA displaces BODIPY®-GDP because KRAS affinity for GTP is greater than its affinity for GDP. The fluorescence intensity decreases as the BODIPY®-GDP is displaced. 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.