Data sheet
| Size | 50 µg, 313 pmol |
| Storage | ‐70°C. Avoid freeze thaw cycles. |
| Application | Recombinant dCas9 (D10A & H840A) protein is suitable for use in imaging of genomic loci in living cells and fixed cells as well as for gene expression regulation. |
| Background | The functions of CRISPR and CRISPR-associated (Cas) genes are essential in adaptive immunity in some bacteria and archaea. CRISPR uses a Cas9 protein to recognize DNA sequences, with target specificity solely determined by a small guide (sg) RNA and a protospacer adjacent motif (PAM). Upon binding to target DNA, the Cas9-sgRNA complex generates a DNA double-stranded break. Based on this RNA- guided nuclease activity, CRISPR has been showed to be a powerful tool in editing the genomes of a broad range of organisms. Furthermore, a repurposed, nuclease-deactivated Cas9 (dCas9) protein has been used to regulate endogenous gene expression and labeling of genomic loci in living and fixed cells. Biotinylated dCas9 was prepared by in vitro biotinylation with biotin to dCas9 at 3:1 ratio. |
| Formulation | Recombinant dCas9 (D10A & H840A) protein expressed in E. coli supplied in a buffer of 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM DTT, 300 mM NaCl, and 50% (v/v) Glycerol. |
| Host/Source | E. coli |
| Sequence | Mutated CRISPR-associated endonuclease Cas9 (amino acids 1 to 1368) with D10A & H840A (Uniprot: Q99ZW2 for Cas9) |
| Amino Acids | D10A/H840A |
| Concentration | 1 µg/ µL, 6.26 µM |
| Physical appearance | Sterile filtered colorless solution. |
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