Key features and details
- Rabbit polyclonal to CRISPR-Cas9
- Suitable for: ICC/IF, IP, Flow Cyt, WB
- Reacts with: Streptococcus pyogenes
- Isotype: IgG
Product nameAnti-CRISPR-Cas9 antibody
See all CRISPR-Cas9 primary antibodies
DescriptionRabbit polyclonal to CRISPR-Cas9
This antibody reacts with CRISPR-Cas9, this antibody has been found to react with 6xHis and Flag tags. If using multiple tags we recommend ab189380.
Tested applicationsSuitable for: ICC/IF, IP, Flow Cyt, WBmore details
Species reactivityReacts with: Streptococcus pyogenes
Recombinant fragment corresponding to Streptococcus pyogenes CRISPR-Cas9 (N terminal). The immunogen was tagged with a 6xHis tag and an S tag this antibody has been found to react with 6xHis and Flag tags.
Database link: Q99ZW2
- WB: HEK-293T cells transfected with DDDDK tag-CRISPR-Cas9 lysate. ICC/IF: U-2 OS cells transfected with DDDDK tag-CRISPR-Cas9. Flow Cyt: HEK-293T cells transfected with DDDDK tag-CRISPR-Cas9. IP: CRISPR-Cas9 IP in U-2 OS cells.
Older lots react with cas9 and the Flag and 6x His tags.
New lots react with only cas9 and the 6x His tag.
Please contact our scientific support team for clarification.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.05% Sodium azide
Concentration information loading...
Our Abpromise guarantee covers the use of ab204448 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||1/1000 - 1/10000.|
|IP||Use at an assay dependent concentration.
Use 10µl per IP.
|Flow Cyt||1/1000 - 1/5000.|
|WB||1/1000 - 1/2000. Predicted molecular weight: 158 kDa.|
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
Lanes 1-2 : Anti-DDDDK tag antibody
Lanes 3-4 : Anti-CRISPR-Cas9 antibody (ab204448) at 1/2000 dilution
Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate
Lanes 2 & 4 : HEK-293T cells transfected with DDDDK tag-CRISPR-Cas9 lysate
Lane 3 : HEK-293T cell lysate
Lysates/proteins at 40 µg per lane.
Predicted band size: 158 kDa
WB: Transfer in nitrocelullose, block: 1 hour 5% milk in TBST (Tween= 0.2%),
Incubation with primary antibody o/n at 4°C in 5% milk TBST (Tween= 0.2%).
Immunofluorescent analysis of U-2 OS (Human bone osteosarcoma epithelial cell line) cells transfected with DDDDK tag-CRISPR-Cas9, labeling CRISPR-Cas9 with ab204448 at 1/10,000 dilution (green). Labeling with anti-DDDDK tag (red). Right hand panel shows merged image with DAPI staining (blue).
Protocol: Fixation with PFA 2% 10 min, permeabilization cold methanol, block with BSA 2% for 1h at RT, incubation with ab204448 1/10,000 1h 30 min at RT.
There is a low background staining with ab204448 which is minimized at 1/10000 dilution.
Flow cytometric analysis of HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cells (upper panel), and transfected with CRISPR-Cas9 (lower panel) labeling CRISPR-Cas9 with ab204448 at 1/5000 dilution.
Cells were fixed with 1% PFA and incubated with the primary antibody (1/5000) at RT for 2 hours.
Western blot analysis of immunoprecipitate using ab204448 in U-2 OS (Human bone osteosarcoma epithelial cell line) cells extracts.
WB: anti-DDDDK tag.
Lane 1: Input.
Lane 2: Immunoprecipitation with control serum (10µl).
Lane 3: Immunoprecipitation with ab204448 (10µl)
ab204448 has been referenced in 9 publications.
- Martin AS et al. A panel of eGFP reporters for single base editing by APOBEC-Cas9 editosome complexes. Sci Rep 9:497 (2019). WB . PubMed: 30679582
- Ajina R et al. SpCas9-expression by tumor cells can cause T cell-dependent tumor rejection in immunocompetent mice. Oncoimmunology 8:e1577127 (2019). WB . PubMed: 31069138
- Gao NJ et al. Functional and Proteomic Analysis of Streptococcus pyogenes Virulence Upon Loss of Its Native Cas9 Nuclease. Front Microbiol 10:1967 (2019). WB . PubMed: 31507572
- St Martin A et al. A fluorescent reporter for quantification and enrichment of DNA editing by APOBEC-Cas9 or cleavage by Cas9 in living cells. Nucleic Acids Res N/A:N/A (2018). WB . PubMed: 29746667
- Woolston BM et al. Rediverting carbon flux in Clostridium ljungdahlii using CRISPR Interference (CRISPRi). Metab Eng N/A:N/A (2018). WB . PubMed: 29906505
- Chaverra-Rodriguez D et al. Targeted delivery of CRISPR-Cas9 ribonucleoprotein into arthropod ovaries for heritable germline gene editing. Nat Commun 9:3008 (2018). IF . PubMed: 30068905
- Wang X et al. Efficient base editing in methylated regions with a human APOBEC3A-Cas9 fusion. Nat Biotechnol N/A:N/A (2018). WB . PubMed: 30125268
- Vojta A et al. Repurposing the CRISPR-Cas9 system for targeted DNA methylation. Nucleic Acids Res N/A:N/A (2016). WB . PubMed: 26969735
- Cinesi C et al. Contracting CAG/CTG repeats using the CRISPR-Cas9 nickase. Nat Commun 7:13272 (2016). WB . PubMed: 27827362