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.

Storage and Stability

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.