General description
CRISPR-associated endonuclease Cas9/Csn1 (UniProt: Q99ZW2; also known as EC:3.1.-.-, SpCas9, SpyCas9) is encoded by the cas9 (also known as csn1, SPy_1046) gene in Streptococcus pyogenes. CRISPR-associated endonuclease Cas9 is an RNA-guided DNA endonuclease enzyme associated with the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) adaptive immunity system in bacteria. Cas9 plays a crucial role in the defense mechanism against against mobile genetic elements (viruses, transposable elements and conjugative plasmids) by cleaving foreign DNA. Cas9 has a bilobed architecture, with a recognition lobe that binds RNA and a nuclease lobe that contains the HNH and RuvC-like nuclease domains. These domains are responsible for cleaving the DNA strands complementary to the guide RNA and the opposite strand, respectively. The enzyme requires a guide RNA that is complementary to the target DNA sequence and a protospacer adjacent motif (PAM) sequence downstream of the target for efficient binding and cleavage. The primary function of Cas9 is to introduce double-stranded breaks at specific locations in the DNA, which are then repaired by the cell′s native repair mechanisms. This feature is harnessed in genome editing to introduce deletions, insertions, or replacements in the DNA sequence. Cas9 application in genome editing has significant therapeutic potential. By targeting and modifying disease-associated genes, Cas9 has been explored as a tool to treat genetic disorders, including sickle cell disease, cystic fibrosis, and certain types of cancer. However, the off-target effects and ethical considerations of genome editing are areas of active investigation and debate. The precise and controlled use of Cas9 remains a critical factor in translating its capabilities into safe and effective therapies.
We are committed to bringing you greener alternative products, which adhere to one or more of the 12 Principles of Green Chemistry. This antibody is preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed, and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information.
ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them.
Immunogen
KLH-conjugated linear peptide corresponding to 24 amino acids from the C-terminal part of human Cas9.
Application
Quality Control Testing
Evaluated by Western Blotting in lysate from Cas9 transfected HEK-293 cells.
Western Blotting Analysis: A 1:10,000 dilution of this antibody detected Cas9 in lysate from HEK-293 cells transfected with Cas9.
Tested Applications
Flow Cytometry Analysis: 1 µg from a representative lot detected Cas9 in one million NIH/3T3 cells transfected with Cas9.
Immunocytochemistry Analysis: A 1:1,000 dilution from a representative lot detected Cas9 in NIH/3T3 cells transfected with Cas9.
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.
Biochem/physiol Actions
Clone 1C12-H1 is a ZooMAb® rabbit recombinant monoclonal antibody that detects Cas9. It targets an epitope within 24 amino acids from the C-terminal region.
Physical form
Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.
Preparation Note
Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.
Reconstitute lyophilized antibody pellet with 25μL of ultrapure water or Phosphate Buffered Saline (PBS). Please refer to our reconstitution protocol and the specific application guidance on the suggested starting dilutions and sample type.
Legal Information
ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany
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.
| biological source | rabbit |
| Quality Segment | 200 |
| recombinant | expressed in HEK 293 cells |
| conjugate | unconjugated |
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| clone | 1C12-H1, recombinant monoclonal |
| description | recombinant, expressed in HEK 293 cells |
| product line | ZooMAb® learn more |
| form | lyophilized |
| mol wt | calculated mol wt 158 kDa,observed mol wt ~158 kDa |
| purified by | using Protein A |
| species reactivity | all |
| packaging | antibody small pack of 25 |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry. |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation |
| sustainability | Greener Alternative Product |
| technique(s) | flow cytometry: suitable,immunocytochemistry: suitable,western blot: suitable |
| isotype | IgGκ |
| epitope sequence | C-terminal half |
| Protein ID accession no. | NP_269215 |
| UniProt accession no. | Q99ZW2 |
| greener alternative category | Aligned |
| shipped in | ambient |
| storage temp. | 2-8°C |
