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Encompassing Amino Acids: 2-186
Application: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: Assay was performed according to the KRAS (G12C) Coupled Nucleotide Exchange Assay Kit (BPS Bioscience #78004) with GDP loaded KRAS (G12C) (BPS Bioscience #100640) and GDP loaded KRAS (G12D) (BPS Bioscience #101312) titrated from 200 nM to 0 nM. Enzyme was incubated with SOS1; positive results show that SOS1 mediated exchange and subsequent RAS-Raf binding is successful.
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 mutation of interest G12D. The protein was affinity purified and loaded with GDP. Unbound GDP was removed by spin column.
Format: Aqueous buffer solution
Formulation: 20 mM HEPES, pH 7.4, 150 mM NaCl, 1 mM DTT
Genbank: NM_033360
Purity: ≥90%
Storage Stability: At least 6 months at -80°C.
Supplied As: Aqueous buffer solution
Tags: N-terminal His-tag
Uniprot: P01116
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
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