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Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: Cas9 activity was measured using DNA Cleavage assay. Active Cas9 enzyme, sgRNA or tracrRNA/crRNA, 5 µl of 4X Cas9 reaction buffer (200 mM Tris-HCl, pH 7.9, 400 mM MgCl2, and 0.4 mg/ml Prionex (w/v 0.04%)) and nuclease-free water were combined to a final reaction volume of 20 µl. The reaction was then incubated at room temperature for 20-60 minutes, followed by the addition of 20-500 ng of target DNA to the Cas9-gRNA complex to initiate the reaction. A subsequent incubation at 37°C for 10-30 minutes. Cleavage of target DNA was confirmed by running a DNA agarose gel.
Description: Recombinant S. pyogenes SpCas9-NLS (conserved hypothetical protein), full length. This construct contains a C-terminal His-Tag. SpCas9-NLS contains a nuclear localization sequence for optimal activity.
Format: Aqueous buffer solution.
Formulation: 50 mM sodium phosphate, pH 7.5, 300 mM NaCl, 1 mM DTT, 150 mM imidazole, and 10% glycerol
Genbank: AAK33936
Purity: ≥70%
Storage Stability: At least 6 months at -80°C.
Tags: C-terminal His-tag
Warnings: Avoid multiple freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
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