The CRISPR associated protein 9 (Cas9) is an RNA-guided DNA nuclease and part of the Streptococcus pyogenes CRISPR antiviral immunity system that provides adaptive immunity against extrachromosomal genetic material (1).
The CRISPR antiviral mechanism of action involves three steps: (i), acquisition of foreign DNA by host bacterium; (ii), synthesis and maturation of CRISPR RNA (crRNA) followed by the formation of RNA-Cas nuclease protein complexes; and (iii), target interference through recognition of foreign DNA by the complex and its cleavage by Cas nuclease activity (2).
The type II CRISPR/Cas antiviral immunity system provides a powerful tool for precise genome editing and has potential for specific gene regulation and therapeutic applications (3).
The Cas9 protein and a guide RNA consisting of a fusion between a crRNA and a trans-activating crRNA (tracrRNA) must be introduced or expressed in a cell.
A 20-nucleotide sequence at the 5' end of the guide RNA directs Cas9 to a specific DNA target site.
As a result, Cas9 can be "programmed" to cut various DNA sites both in vitro and in cells and organisms.
CRISPR/Cas9 genome editing tools have been used in many organisms, including mouse and human cells (4,5).
Research studies demonstrate that CRISPR can be used to generate mutant alleles or reporter genes in rodents and primate embryonic stem cells (6-8).
Cas9 (S.
aureus) is a Cas9 ortholog that is smaller, but as efficient, as the more commonly used Cas9 ortholog, Cas9 (S.
pyogenes) (9).
Applications:Suitable for use in Immunofluorescence, Immunocytochemistry, Immunoprecipitation and Western Blot.
Other applications not tested.
Recommended Dilutions:Western Blot: 1:1000Immunoprecipitation: 1:200Immunofluorescence (IC): 1:400-1:1600Optimal dilutions to be determined by the researcher.
Storage and Stability:May be stored at 4℃ for short-term only.
Aliquot to avoid repeated freezing and thawing.
Store at -20℃.
Aliquots are stable for 12 months after receipt.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
仕様
Size：100ul
抗体種類：Mab
宿主：mouse
Grade：Purified
Purity：Purified
Form：Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/mL BSA, <0.02% sodium azide, 50% glycerol.
Specificity：Recognizes endogenous levels of total Cas9 (S. aureus) protein. Does not cross-react with Cas9 (S. pyogenes), AsCpf1 (Strain BV3L6), and FnCpf1 (Strain U112) proteins.
Clone # USB：6H4
Isotype：IgG2b
Calc Applications Abbrev：IC IF IP WB
Immunogen：Recombinant protein corresponding to the N-terminus of S. aureus Cas9 protein.
