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General description
CRISPR-associated endonuclease Cas9/Csn1 (UniProt Q99ZW2; also known as SpyCas9) is encoded by the Streptococcus pyogenes serotype M1 gene Cas9 (also known as csn1) gene (Gene ID 901176). Clustered regularly interspaced short palindromic repeats (CRISPRs) are DNA loci containing short repetitions of base sequences complementary to antecedent mobile elements and target invading nucleic acids. Each repetition is followed by short segments of spacer DNA from previous exposures to a virus. Cas9 is an essential endonuclease in Streptococcus pyogenes serotype M1′s CRISPR immune system that confers resistance to foreign genetic elements such as plasmids and phages. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). In type II CRISPR systems, correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc), and Cas9. Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer. The target strand not complementary to crRNA is first cut endonucleolytically, and then trimmed 3′-5′ exonucleolytically.
~160 kDa observed. 158.4 kDa calculated. Uncharacterized bands may be observed in some lysate(s).
Immunogen
His-tagged recombinant Cas9 C-terminal fragment excluding the active sites RuvC-II & RuvC-III.
Application
Anti-Cas9, CT, clone 6G12-H11, Cat. No. MABE1668, is a highly specific mouse monoclonal antibody that targets CRISPR-associated endonuclease Cas9/Csn1 and has been tested in Immunocytochemistry and Western Blotting.
Immunocytochemistry Analysis: A 1:50 dilution from a representative lot detected exogenously expressed full-length nuclease-dead Cas9 in transfected NIH/3T3 cells fixed with 4% paraformaldehyde and permeabilized by 0.3% Triton X-100.
Research Category
Epigenetics & Nuclear Function
Biochem/physiol Actions
Clone 6G12-H11 targets an epitope within Cas9 C-terminal region outside the active sites RuvC-II & RuvC-III.
Physical form
Format: Purified
Protein G purified.
Purified mouse IgG1κ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Preparation Note
Stable for 1 year at 2-8°C from date of receipt.
Analysis Note
Evaluated by Western Blotting in NIH/3T3 cells.
Western Blotting Analysis: 0.25 µg/mL of this antibody detected exogenously expressed full-length nuclease-dead Cas9 in 10 µg of lysate from transfected NIH/3T3 cells.
Other Notes
Concentration: Please refer to lot specific datasheet.
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 | mouse |
| Quality Segment | 100 |
| conjugate | unconjugated |
| antibody form | purified immunoglobulin |
| antibody product type | primary antibodies |
| clone | 6G12-H11, monoclonal |
| species reactivity | Streptococcus pyogenes |
| species reactivity (predicted by homology) | all |
| technique(s) | immunocytochemistry: suitable,western blot: suitable |
| isotype | IgG1κ |
| NCBI accession no. | NP_269215 |
| UniProt accession no. | Q99ZW2 |
| shipped in | ambient |
| target post-translational modification | unmodified |
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