General description
CRISPR-associated endonuclease Cas9 (UniProt: J7RUA5; also known as SaCas9, Cas9) is encoded by the Cas9 gene in Streptococcus aureus. Cas9 belongs to subtype II-A subfamily with one HNH Cas9-type domain. 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. Cas9 has a bi-lobed architecture with a recognition lobe (residues 41-425) and a discontinuous nuclease lobe (NUC, residues 1-40 and 453-1053); the crRNA-target DNA lies in a channel between the 2 lobes. The NUC lobe has 2 endonuclease domains. The discontinuous RuvC-like domain in NUC cleaves the target DNA non-complementary to crRNA while the HNH nuclease domain cleaves the target DNA complementary to crRNA. Cas9 serves as a RNA-guided DNA endonuclease that cleaves double-stranded DNA targets with a protospacer adjacent motif (PAM) and complementarity to the guide RNA. The homologous region of the Streptococcus pyogenes is not closely related in amino acid sequence in S. aureus, hence this antibody does not recognize that protein.
~55 kDa observed. Uncharacterized bands may be observed in some lysate(s).
Application
Anti-Cas9 (Rabbit Polyclonal), Cat. No. AB356480, is a highly specific rabbit polyclonal antibody that targets CRISPR-associated endonuclease Cas9 and has been tested in Immunofluorescence and Western Blotting.
Immunofluorescence Analysis: A 1:1,000 dilution from a representative lot detected Cas9 in HEK293 cells with transfected with GFP-Cas9-SA (Courtesy of Dr. Gerry Shaw).
Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected Cas9 in HEK293 cells overexpressing fusion protein GFP-Cas9 (Courtesy of Dr. Gerry Shaw).
Biochem/physiol Actions
This rabbit polyclonal antibody detects Cas9 from Staphylococcus aureus. It targets an epitope within 251 amino acids from the C-terminal end. It does not react with Cas9 from Streptococcus pyogenes.
Analysis Note
Evaluated by Western Blotting in HEK293 cells overexpressing fusion protein GFP-Cas9.
Western Blotting Analysis: 0.2 µg/mL of this antibody detected Cas9 (Rabbit Polyclonal) in 40 µg of HEK293 cells overexpressing fusion protein GFP-Cas9 from Staphylococcus aureus.
Other Notes
Concentration: Please refer to lot specific datasheet.
| biological source | rabbit |
| Quality Segment | 100 |
| conjugate | unconjugated |
| antibody form | serum |
| antibody product type | primary antibodies |
| clone | polyclonal |
| species reactivity | Staphylococcus aureus, bacteria |
| packaging | antibody small pack of 25 μL |
| technique(s) | immunofluorescence: suitable,western blot: suitable |
| isotype | IgG |
| UniProt accession no. | J7RUA5 |
| shipped in | ambient |
| target post-translational modification | unmodified |
