KRAS (G12C), Isoform A, BODIPY-GDP Loaded, His-Tag
Catalog #
100537
$395*
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Description
Recombinant human KRAS (GTPase), isoform a, encompassing amino acids 2-186 (end). In this construct the N-terminal end of the protein contains a His-tag (6xHis) followed by an optimal TEV protease target sequence that allows cleavage of the His-tag from the protein (TEV: tobacco etch virus cysteine protease, BPS Bioscience #50308). The protein also contains cancer-associated mutation G12C. KRAS (G12C) His-Tag (BPS Bioscience #100413) was affinity purified and was loaded with BODIPY-GDP. Unbound BODIPY-GDP was removed by spin column and the protein is ready for a GDP/GTP exchange assay.
BODIPY™ FL GDP ((ThermoFisher Scientific, Guanosine 5'-Diphosphate, BODIPY™ FL 2'-(or-3')-O-(N-(2-Aminoethyl) Urethane), Bis (Triethylammonium) Salt), 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. Invitrogen BODIPY® FL dye is a green-fluorescent dye with similar excitation and emission to fluorescein (FITC) or Alexa Fluor 488. It is characterized by a high extinction coefficient and high quantum yield and is relatively insensitive to pH changes. Its hydrophobic properties make it useful with lipophilic compounds. The dye has an excited-state lifetime of 5 nanoseconds or longer, making it useful for fluorescence polarization–based assays.
Storage/Stability
At least 6 months at –80°C.
Assay Conditions
Assay was performed according to the KRAS(G12C) Nucleotide Exchange Assay Kit (BPS Bioscience #79859) with BODIPY-GDP-loaded KRAS(G12C) concentrated at 5 µM (~ 0.12 mg/ml) and GTP titrated 3-fold from 100 µM – 0 µM.
Instructions for Use
Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.
Applications
Useful for KRAS(G12C) Nucleotide Exchange Assay Kit (BPS Bioscience #79859), inhibitors screening, and selectivity profiling.
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