KRAS (G12D), Isoform A, His-Tag, BODIPY-GDP Loaded Recombinant

• 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 proteaseBPS 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

• UniProt:

  P01116

• Tag:

  N-terminal His-tag

• Label:

  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

• Applications:

  Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

• Purity:

  ≥ 90%

• Format:

  Aqueous buffer solution

• Shipping Conditions:

  -80°C

• Storage Conditions:

  At least 6 months at -80°C.

• Calculated Molecular Weight:

  23 kDa

• Formulation:

  20 mM HEPES, pH 7.4,150 mM NaCl and 1 mM DTT