This product is prepared by adding 50 µL of the reconstitution solution to the 250 µg package or 30 µL of the reconstitution solution to the 50 µg. The reconstitution solution is 50% glycerol in water. A prepared stock solution may be stored at -20°C. The dilution buffer is 20 mM Na-Hepes pH 7.5, 200 with mM NaCl. Please see the link below to review the full product datasheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/138/442/cas9d10aprbul.pdf
CAS9D10APR
Cas9-D10A Nickase Protein
from Streptococcus pyogenes with D10A mutation, recombinant, expressed in E. coli, 1X NLS
Synonym(s):
Cas9 Nickase, Cas9D10A, Nickase Cas9
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50 μG
$97.18
250 μG
$354.32
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50 μG
$97.18
250 μG
$354.32
About This Item
UNSPSC Code:
12352202
NACRES:
NA.51
Recommended Products
Quality Level
assay
≥95% (SDS-PAGE)
form
lyophilized powder
packaging
pkg of 1 kit (3 components)
shipped in
dry ice
storage temp.
−20°C
Related Categories
General description
Cas9 (CRISPR (clustered regularly interspaced short palindromic repeats)-associated protein-9 nuclease) nickase creates double-stranded breaks through two nuclease domains, named RuvC and HNH.[1]
Recombinant Cas9-D10A Nickase protein from Streptococcus pyogenes (~160 KD) is a ready-to-use reagent for genome engineering experiments. When combined with target-specific paired guide RNAs, S. pyogenes Cas9-D10A nickase will act as a targeted nuclease suitable for transfection of cell cultures and for the accelerated development of genetically-modified animals via one-cell embryo injection.
Application
Functional Genomics/Target Validation/Genome Editing
Features and Benefits
- Highly specific
- Highly active
- Ready-to-inject/transfect
Packaging
pkg of 50 μg (≥ 300 pmol)
pkg of 250 μg (≥ 1500 pmol)
pkg of 250 μg (≥ 1500 pmol)
Components
Each kit consists of:
- one vial of Cas9-D10A Nickase recombinant protein
- one vial containing 1 mL of 1× dilution buffer
- one vial containing 1 mL of nuclease-free water with glycerol
Principle
CRISPR/Cas systems are employed by bacteria and archaea as a defense against invading viruses and plasmids. Recently, the type II CRISPR/Cas system from the bacterium Streptococcus pyogenes has been engineered to function in eukaryotic systems using two molecular components: a single Cas9 protein and a non-coding guide RNA (gRNA). The Cas9 endonuclease can be programmed with a gRNA, directing a DNA double-strand break (DSB) at a desired genomic location. Similar to DSBs induced by zinc finger nucleases (ZFNs), the cell then activates endogenous DNA repair processes, either non-homologous end joining (NHEJ) or homology-directed repair (HDR), to heal the targeted DSB.
Reconstitution
Lyophilized S. pyogenes Cas9-D10A Nickase protein should be resuspended in the Reconstitution solution provided to desired concentration. Gently tap tube to completely dissolve lyophilized powder, incubate for 10 minutes on ice, and spin tube to bring material to bottom of tube.
Other Notes
Use our CRISPR Selection Tool to order gRNA
Check out our other MISSION® Cas9 Proteins at SigmaAldrich.com/CRISPRproteins
See the product in our recent article on bioRxiv Highly efficient and specific genome editing in human cells with paired CRISPR-Cas9 nickase ribonucleoproteins
Check out our other MISSION® Cas9 Proteins at SigmaAldrich.com/CRISPRproteins
See the product in our recent article on bioRxiv Highly efficient and specific genome editing in human cells with paired CRISPR-Cas9 nickase ribonucleoproteins
Legal Information
MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany
Storage Class
11 - Combustible Solids
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