Product nameAnti-CRISPR-Cas9 antibody [7A9-3A3]
See all CRISPR-Cas9 primary antibodies
DescriptionMouse monoclonal [7A9-3A3] to CRISPR-Cas9
Tested applicationsSuitable for: ICC/IF, IHC - Wholemount, WBmore details
Species reactivityReacts with: Streptococcus pyogenes
Recombinant fragment corresponding to Streptococcus pyogenes CRISPR-Cas9 (N terminal).
- WB: S2 cells transfected with CRISPR-Cas9 ICC-IF: NIH/3T3-Cas9 transfected cells.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term.
Storage bufferpH: 7.4
Preservative: 0.02% Sodium azide
Some batches contain 6.97% L-Arginine as a stabilizing agent. For lot-specific buffer information, please contact our Scientific Support team.
Concentration information loading...
PurityProtein G purified
Light chain typekappa
Our Abpromise guarantee covers the use of ab191468 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 - 10 µg/ml.|
|IHC - Wholemount||1/500. PubMed: 27638686|
|WB||Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 160 kDa (predicted molecular weight: 160 kDa).
We recommend using 3% milk as the blocking agent for Western blot.
Relevance[FUNCTION] CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids). CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA) (Probable). In type II CRISPR systems correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc) and this protein. The tracrRNA serves as a guide for ribonuclease 3-aided processing of pre-crRNA. Subsequently Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer. The target strand not complementary to crRNA is first cut endonucleolytically, then trimmed by 3'-5' exonucleolytically. DNA-binding requires protein and both RNA species. Cas9 probably recognizes a short motif in the CRISPR repeat sequences (the PAM or protospacer adjacent motif) to help distinguish self versus nonself.
- Cas9 antibody
- CRISPR-associated endonuclease Cas9/Csn1 antibody
- CRISPR-Cas9/Csn1 antibody
All lanes : Anti-CRISPR-Cas9 antibody [7A9-3A3] (ab191468) at 5 µg/ml
Lane 1 : S2 non-transfected cell lysate
Lane 2 : S2 cells transfected with CRISPR-Cas9 plasmid
Lysates/proteins at 20 µg per lane.
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 160 kDa
Observed band size: 160 kDa
Exposure time: 4 minutes
We recommend using 3% milk as the blocking agent in Western Blot.
ab191468 stained in NIH3T3 cells. Untreated and Cas9 transfected cells were fixed with 4% paraformaldehyde (10min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab191468 at 10µg/ml and ab6046 (Rabbit polyclonal to beta tubulin) at 1ug/ml overnight at +4°C. The secondary antibodies were ab150117 (colored green) used at 1 ug/ml and ab150087 (pseudo-colored red) used at 2ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.
This product has been referenced in:
- Park H et al. In vivo neuronal gene editing via CRISPR-Cas9 amphiphilic nanocomplexes alleviates deficits in mouse models of Alzheimer's disease. Nat Neurosci 22:524-528 (2019). Read more (PubMed: 30858603) »
- Simhadri VL et al. Prevalence of Pre-existing Antibodies to CRISPR-Associated Nuclease Cas9 in the USA Population. Mol Ther Methods Clin Dev 10:105-112 (2018). Read more (PubMed: 30073181) »