Anti-CRISPR/CAS9 -FITC antibody, Mouse monoclonal
SIGMA/SAB4200735 - clone 7A9-3A3, purified from hybridoma cell culture
Synonym: Anti-
Product Type: Chemical
Immunofluorescence Human HEK-293T over-expressing CRISPR/CAS9 cells were fixed and permeabilized with 4% paraformaldehyde followed by 0.5% Triton X-100. Fixed cells were stained with 1:400 Anti-CRISPR/CAS9-FITC, Mouse monoclonal, Clone: 7A9-3A3 (Cat. No. SAB4200735). Cells were counterstained with DAPI (blue) to stain nuclei.
| antibody form | purified from hybridoma cell culture |
| antibody product type | primary antibodies |
| clone | 7A9-3A3, monoclonal |
| concentration | ~1.0 mg/mL |
| conjugate | FITC conjugate |
| form | buffered aqueous solution |
| isotype | IgG1 |
| Quality Level | 200  |
| shipped in | dry ice |
| storage condition | protect from light |
| storage temp. | −20°C |
| target post-translational modification | unmodified |
| technique(s) | immunofluorescence: 1:200-1:400 using human HEK-293T cells over-expressing CAS9 protein |
| UniProt accession no. | Q99ZW2  |
| Application: | Monoclonal Anti-CRISPR/CAS9-FITC recognizes the CAS9 protein in CAS9 construct over-expression systems. The antibody may be used in various immunochemical techniques including Immunofluorescence. The antibody successfully recognizes mutant Cas9 variants, including nickase Cas9 and dead Cas9 (dCas9) and does not cross react with FnCas9 from Francisella novicida bacteria and Cpf1 proteins from Acidaminococcus sp. (strain BV3L6) and Lachnospiraceae bacterium ND2006. |
| General description: | CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) belongs to the type II CRISPR/CAS9 system. It is part of an adaptive immune system of the Streptococcus pyogenes SF370, protecting from pathogens′ target genes by cleaving the foreign DNA in a sequence-dependent manner. The type II CRISPR/Cas system has been adapted to expression in eukaryotic cells. The Cas9 endonuclease can be engineered with a single gRNA (guide RNA), directing a DNA double-strand break (DSB) at a desired genomic location. As a result, the cell activates endogenous DNA repair processes, either non-homologous end joining (NHEJ) or homology-directed repair (HDR). In comparison to other genome-editing technologies such as designer zinc fingers (ZFs), transcription activator–like effectors (TALEs) and homing meganucleases, the CRISPR/CAS9 system is a scalable, affordable and easy to engineer. Therefore, the anti-CRISPR/CAS9-FITC conjugated antibody can be a useful tool for detecting CRISPR/CAS9 positively transfected cells, revealing DSB sites in their genome. |
| Immunogen: | Recombinant protein within the N-terminal region of Streptococcus pyogene Cas9 |
| Other Notes: | This product is for R&D use only, not for drug, household, or other uses. In order to obtain best results in different techniques and preparations we recommend determining optimal working concentration by titration test. |
| Physical form: | Solution in 0.01M phosphate buffered saline pH 7.4 containing 1% bovine serum albumin and 15mM sodium azide. |
| Preparation Note: | For continuous use, store at 2-8°C, protected from light for up to one month. For extended storage, freeze in working aliquots. Repeated freezing and thawing is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilution samples should be discarded if not used within 12 hours. |
| RIDADR | NONH for all modes of transport |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12352203 |