The recommended 1X reaction buffer is 20 mM Tris, pH 7.4, 100 mM NaCl, 5 mM MgCl2, 1 mM DTT. The dilution buffer, which is included in this kit, is recommended for further dilution of reconstituted proteins, if required.
CAS9PL
Cas9 Plus Protein
from Streptococcus pyogenes, recombinant, expressed in E. coli, 3X NLS
Synonym(s):
Cas9, SpCas9, SpyCas9
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| Size/SKU | Availability | Price |
|---|---|---|
50 μg | Check Cart for Availability | $184.00 |
250 μg | Check Cart for Availability | $682.00 |
About This Item
NACRES:
NA.51
UNSPSC Code:
12352200
Form:
lyophilized powder
Assay:
≥95% (SDS-PAGE)
Recombinant:
expressed in E. coli
recombinant
expressed in E. coli
Quality Segment
assay
≥95% (SDS-PAGE)
form
lyophilized powder
packaging
pkg of 1 kit (4 components)
application(s)
CRISPR
CRISPR
shipped in
wet ice
storage temp.
−20°C
General description
Recombinant Cas9 Plus protein from Streptococcus pyogenes (~160 KD) is a ready-to-use reagent for genome engineering experiments. When combined with target-specific guide RNAs, Streptococcus pyogenes Cas9 Plus protein 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
Biochem/physiol Actions
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.
Features and Benefits
- Enhanced specificity compared to wild type Cas9
- Increased activity compared to enhanced specificity Cas9
- Ready to inject/transfect
Packaging
pkg of 50 μg (≥ 300 pmol)
pkg of 250 μg (≥ 1500 pmol)
pkg of 250 μg (≥ 1500 pmol)
Preparation Note
Lyophilized S. pyogenes Cas9 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
Each kit consists of:
- one vial of Cas9 Plus Recombinant Protein
- one vial of Plus Transfection Buffer
- one vial of Reconstitution Buffer
- one vial of Dilution Buffer
Use our CRISPR Selection Tool to order gRNA
Check out our other MISSION® Cas9 Proteins at SigmaAldrich.com/CRISPRproteins
Check out our other MISSION® Cas9 Proteins at SigmaAldrich.com/CRISPRproteins
Legal Information
MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany
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This Item | |||
|---|---|---|---|
| description from Streptococcus pyogenes, recombinant, expressed in E. coli, 3X NLS | description from Streptococcus pyogenes, recombinant, expressed in E. coli, 1X NLS | description from Streptococcus pyogenes, recombinant, expressed in E. coli, 3X NLS | description - |
| recombinant expressed in E. coli | recombinant expressed in E. coli | recombinant expressed in E. coli | recombinant expressed in E. coli |
| assay ≥95% (SDS-PAGE) | assay ≥95% (SDS-PAGE) | assay ≥95% (SDS-PAGE) | assay - |
| form lyophilized powder | form lyophilized powder | form lyophilized powder | form liquid |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. −20°C |
| shipped in wet ice | shipped in wet ice | shipped in wet ice | shipped in dry ice |
| Quality Level 200 | Quality Level 300 | Quality Level 300 | Quality Level - |
WGK
WGK 3
Storage Class
10 - Combustible liquids
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Hi, which reaction buffer can I use for Cas9 Plus-sgRNA complex for in vitro assay? I need to check cleavage of DNA in vitro before injections into animals and I didnt find any protocols or reccomendations in datasheets. Can I use dilution buffer?
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