Description General description CRISPR-associated endonuclease Cas9/Csn1 (UniProt: Q99ZW2; also known as SpCas9, SpyCas9) is encoded by the Cas9 (also known as Csn1) gene (Gene ID: 901176) in Streptococcus pyogenes. 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 Staphylococcus aureus is not closely related in amino acid sequence in Streptococcus pyogenes, hence this antibody does not recognize that protein. Specificity This rabbit polyclonal antibody detects Cas9 in Streptococcus pyogenes. It targets an epitope within 18 amino acids from the C-terminal region. This antibody does not react with Cas9 from Staphylococcus aureus. Immunogen Epitope: C-terminus KLH-conjugated linenar peptide corresponding to 18 amino acids from the C-terminal region of Streptococcus pyogenes. Application Anti-Cas9 Antibody, Cat. No. ABS2202, is a highly specific rabbit polyclonal antibody that targets Cas9 and has been tested for use in Immunocytochemistry and Western Blotting. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected Cas9 in HEK293 cells transfected with Cas9 from Streptococcus pyogenes. Immunocytochemistry Analysis: 1 µg/mL from a representative lot detected Cas9 in non-transfected HEK293H and HEK293H transfected with pCDNA3.0-Cas9. Western Blotting Analysis: 10 µg/mL from a representative lot detected Cas9 in HEK293H cell lysate from cells transfected with pCDNA3.0-Cas9. Research Category Epigenetics & Nuclear Function Target description 158.44 kDa calculated. Physical form Affinity Purified Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide. Storage and Stability Stable for 1 year at 2-8°C from date of receipt. Disclaimer Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. Quality Evaluated by Western Blotting in HEK293 cells transfected with Cas9 from Streptococcus pyogenes. Western Blotting Analysis: A 1:500 dilution of this antibody detected Cas9 in HEK293 cells transfected with Cas9 from Streptococcus pyogenes. Other Notes Concentration: Please refer to lot specific datasheet.