This product features a 3X FLAG-biotin tag and does not include a histidine tag.
DCAS9PROT
dCas9-3XFLAG™-Biotin Protein
from Streptococcus pyogenes with D10A and H840A mutations, recombinant, expressed in E. coli, 1X NLS
Synonym(s):
Cas9 D10A H840A, Dead Cas9, dCas9, catalytically inactive Cas9
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About This Item
NACRES:
NA.51
UNSPSC Code:
12352202
Form:
lyophilized powder
Assay:
≥90% (SDS-PAGE)
Recombinant:
expressed in E. coli
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recombinant
expressed in E. coli
Quality Level
assay
≥90% (SDS-PAGE)
form
lyophilized powder
packaging
pkg of 1 kit (3 components)
shipped in
ambient
storage temp.
−20°C
Related Categories
1 of 4
This Item | |||
|---|---|---|---|
| recombinant expressed in E. coli | recombinant expressed in E. coli | recombinant expressed in E. coli | recombinant expressed in E. coli |
| assay ≥90% (SDS-PAGE) | assay ≥95% (SDS-PAGE) | assay ≥90% (SDS-PAGE) | assay ≥95% (SDS-PAGE) |
| form lyophilized powder | form lyophilized powder | form lyophilized powder | form lyophilized powder |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. −20°C |
| shipped in ambient | shipped in wet ice | shipped in wet ice | shipped in ambient |
| packaging pkg of 1 kit (3 components) | packaging pkg of 1 kit (3 components) | packaging pkg of 1 kit (3 components) | packaging pkg of 1 kit (3 components) |
General description
Recombinant dCas9-3X FLAG-Biotin protein from Streptococcus pyogenes (~165 KD) is a ready-to-use reagent for genome engineering and genomic DNA detection, isolation and imaging experiments. When combined with target-specific guide RNAs, dead Streptococcus pyogenes Cas9 protein acts as a DNA targeting protein suitable for transfection of cell cultures, microinjection, DNA pull-down, chromatin enrichment, microbial identification and in situ DNA imaging. At the C terminus, the protein is tagged with a SV40 large T antigen nuclear localization signal (NLS) for nuclear import and both a 3XFLAG epitope and biotin for anti-FLAG®-based or streptavidin-based experiments.
Application
Functional Genomics/Target Validation/Genome Editing/Molecular Biology/Synthetic Biology/DNA detection/isolation/DNA imaging
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 rendered inactive (dCas9) with mutations to the two protein domains, RuvC and HnH (D10A and H840A respectively), which are responsible for nuclease activity. The protein can then be programmed with a gRNA and directed to bind at a desired sequence of DNA.
Features and Benefits
- Highly specific
- Suitable for genomic DNA detection/isolation
- Ready-to-inject/transfect
Packaging
- pkg of 50 μg (≥ 300 pmol)
- pkg of 250 μg (≥ 1500pmol)
Preparation Note
Lyophilized dCas9 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 dCas9 recombinant protein
- one vial containing 1 mL of 1× dilution buffer
- one vial containing 1 mL of Nuclease free water with glycerol
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
3xFLAG is a trademark of Sigma-Aldrich Co. LLC
FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany
Storage Class
10 - Combustible liquids
wgk
WGK 3
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Matthew D Newton et al.
Methods in molecular biology (Clifton, N.J.), 2478, 349-378 (2022-09-06)
The discovery of CRISPR/Cas9 as an easily programmable endonuclease heralds a new era of genetic manipulation. With this comes the prospect of novel gene therapy approaches, and the potential to cure previously untreatable genetic diseases. However, reports of spurious off-target
Protocols
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Global Trade Item Number
| SKU | GTIN |
|---|---|
| DCAS9PROT-250UG | 04061835240401 |
| DCAS9PROT-50UG | 04061835240395 |
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Is this construct 6xHis Tagged?
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Is the biotin in Product No. DCAS9PROT attached to the protein by in vivo BirA biotinylation or chemical attachment?
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The biotin in Product No. DCAS9PROT is attached to the protein through in vivo BirA biotinylation.
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