KRAS (G12D), Isoform A, His-Tag, BODIPY-GDP Loaded Recombinant
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 mutation of interest G12D. The protein 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
≥90%
Aqueous buffer solution
20 mM HEPES, pH 7.4, 150 mM NaCl and 1 mM DTT
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
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