Product nameAnti-CRISPR-Cas9 antibody [7A9-3A3] (HRP)
See all CRISPR-Cas9 primary antibodies
DescriptionMouse monoclonal [7A9-3A3] to CRISPR-Cas9 (HRP)
Tested applicationsSuitable for: WBmore details
Species reactivityReacts with: Streptococcus pyogenes
Recombinant fragment corresponding to Streptococcus pyogenes CRISPR-Cas9 (N terminal).
- WB: S2 cells transfected with CRISPR-Cas9
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. Store In the Dark.
Storage bufferpH: 7.40
Preservative: 0.1% Proclin
Constituents: PBS, 1% BSA, 30% Glycerol
Concentration information loading...
PurityProtein G purified
Light chain typekappa
Our Abpromise guarantee covers the use of ab202580 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/5000. Detects a band of approximately 180 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] (HRP) (ab202580) at 1/5000 dilution
Lane 1 : Cas 9 negative control (S2 non-transfected cell lysate)
Lane 2 : Cas 9 transfected cells (S2 cells transfected with CRISPR-Cas9 lysate)
Lysates/proteins at 20 µg per lane.
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 160 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted?
Exposure time: 2 minutes
This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab202580 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.
This product has been referenced in:
- Koch B et al. Generation and validation of homozygous fluorescent knock-in cells using CRISPR-Cas9 genome editing. Nat Protoc 13:1465-1487 (2018). Read more (PubMed: 29844520) »
- Zhang S & Voigt CA Engineered dCas9 with reduced toxicity in bacteria: implications for genetic circuit design. Nucleic Acids Res 46:11115-11125 (2018). Read more (PubMed: 30289463) »