78355

KRAS(G12D) Nucleotide Exchange Assay Kit

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.

5067.68 €

78519

KRAS(G12V) Nucleotide Exchange Assay Kit

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.

4620.75 €

78565

KRAS(G12C) Coupled Nucleotide Exchange Assay Kit

The KRAS(G12C) Coupled Nucleotide Exchange Assay Kit is designed for screening and profiling of KRAS(G12C) antagonists/inhibitors by monitoring the binding of an effector protein such as the Ras binding domain of Raf1, (RBD-cRaf) to KRAS(G12C). With this kit, a few simple steps on a microtiter plate are required for nucleotide exchange detection. 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.SOS1 (son of sevenless) is a guanine nucleotide exchange factor that facilitates the exchange of GDP for GTP. GDP-loaded KRAS(G12C) is in an inactive state, and does not interact with the Ras-binding domain (RBD) of cRAF.  SOS1 assists in the release of GDP from KRAS(G12C) so that GTP can occupy the nucleotide binding pocket. This results in a conformational change in KRAS(G12C) that permits its binding to RBD-cRAF. The KRAS(G12C) Coupled Nucleotide Exchange Assay Kit utilizes GST-tagged RBD-cRAF and His-tagged KRAS (G12C) to assay binding of KRAS(G12C) to RBD-cRAF in the Alpha assay. Glutathione acceptor and Ni chelate donor beads are brought into proximal range by the binding of KRAS(G12C) and RBD-cRAF, enabling the energy transfer from the donor to acceptor beads after laser excitation. 

2204.32 €

GA-E0662HM-96Tests

Human Guanine nucleotide Exchange Factor(GEF)ELISA Kit

719.12 €

MBS265644-10x96StripWells

Mouse Guanine nucleotide Exchange Factor (GEF) ELISA Kit

4423.55 €

MBS265644-48StripWells

Mouse Guanine nucleotide Exchange Factor (GEF) ELISA Kit

566.73 €