Anti-Cas9 Rabbit Monoclonal Antibody for WB, IF/ICC, ELISA - Q99ZW2

Core Product Specifications and Parameters

Biological Background: Cas9 Function and Localization

• Cas9 (CRISPR-associated endonuclease Cas9/Csn1) is a 158.4 kDa RNA-guided DNA endonuclease that forms the enzymatic core of the type II CRISPR-Cas adaptive immune system in Streptococcus pyogenes. Related references: PMID:21455174
• In its native biological role, Cas9 protects bacteria against invading plasmids and viral DNA by incorporating short foreign sequences (spacers) into CRISPR loci, providing a genetic memory for sequence-specific immunity. Related references: PMID:21455174
• Cas9 requires two small RNAs—a CRISPR RNA (crRNA) and a trans-activating crRNA (tracrRNA)—for both target DNA binding and nuclease activity; it is catalytically inactive in the absence of these guide RNAs. Related references: PMID:24270795 PMID:24476820
• Target recognition is governed by a short protospacer adjacent motif (PAM; 5'-NGG-3') within the CRISPR repeat, which serves to discriminate self from non-self DNA; PAM binding initiates DNA strand separation and heteroduplex formation. Related references: PMID:24476820
• Upon PAM recognition, Cas9 catalyzes endonucleolytic cleavage of the target strand non-complementary to the crRNA, followed by 3'-5' exonucleolytic trimming; both nuclease steps are dependent on magnesium or manganese ions. Related references: PMID:24476820
• In CRISPR RNA maturation, Cas9 does not catalyze processing; it stabilizes the pre-crRNA:tracrRNA duplex, enabling accurate cleavage by host ribonuclease III. Related references: PMID:24270795

Experimental Guidance and Technical Tips

• The immunogen spans amino acids 450-730 of Cas9, a region encompassing the HNH nuclease domain and the PAM-interacting domain. For western blotting, consider using lysates from Cas9-transfected cells or purified recombinant full-length Cas9 as a positive control; the predicted molecular weight is 158 kDa.
• In immunofluorescence/ICC, Cas9 localization may vary depending on guide RNA and target. Validate fixation and permeabilization conditions with appropriate controls, and confirm specificity using Cas9-knockout or knock-down cell lines if available.
• For ELISA, the recombinant Cas9 fragment can be used to establish a standard curve; however, assess cross-reactivity with endogenous Cas9 or other family members in the relevant biological matrix before quantitative application.
• This antibody is predicted to cross-react widely, including with human Cas9. When using samples from other species, initial validation is recommended to confirm expected reactivity.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio provides diagnostic manufacturers, research laboratories, and biotechnology companies with one-stop access to high-quality IVD raw materials, technical services, and consulting—supporting projects from concept to clinic. This Cas9 monoclonal antibody is tailored for CRISPR-based research, gene editing tool development, and diagnostic assay design. We offer validated antibody pairs, optimized monoclonal and polyclonal antibodies, bulk ancillary reagents, and rare-target raw-material sourcing to streamline your workflow.