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.
Specificity
Clone 10C11-A12 recognizes an epitope located in the C-terminal region of Cas9.
Immunogen
His-tagged recombinant S.pyogenes Cas9 C-terminal fragment.
Application
Anti-Cas9 Antibody, C-term. clone 10C11-A12 is an antibody against Cas9 for use in Immunocytochemistry, Immunoprecipitation, and Western Blotting.
Immunocytochemistry Analysis: 33.5 µg/mL from a representative lot detected doxycycline-induced expression of exogenously transfected S. pyogenes Cas9 construct under the control of the PTight (Tet-ON) promoter by fluorescent immunocytochemistry staining of methanol-fixed, acetone-permeabilized HeLa transfectants (Courtesy of Dr. Stefan Schuechner and Dr. Egon Ogris, Medical University of Vienna, Austria).
Immunoprecipitation Analysis: 13.4 µg from a representative lot immunoprecipitated exogenously expressed Cas9 in 250 µg of lysate from transfected, but not untransfected, HeLa cells (Courtesy of Dr. Stefan Schuechner and Dr. Egon Ogris, Medical University of Vienna, Austria).
Western Blotting Analysis: 1.675 µg/mL from a representative lot detected exogenously expressed Cas9 in 20 µg of lysate from transfected, but not untransfected, HeLa cells (Courtesy of Dr. Stefan Schuechner and Dr. Egon Ogris, Medical University of Vienna, Austria).
Western Blotting Analysis: Clone 10C11-A12 hybridoma culture supernatant detected doxycycline treatment-induced expression of a FLAG-tagged Cas9 construct under the control of the PTight promoter in a HeLa-derived Tet-ON cell line (Courtesy of Dr. Stefan Schuechner and Dr. Egon Ogris, Medical University of Vienna, Austria).
Quality
Evaluated by Western Blotting in Cas9-transfected HeLa cell lysate.
Western Blotting Analysis: 0.5 µg/mL of this antibody detected the exogenously expressed Cas9 in 10 µg lysate from transfected HeLa cells.
Target description
~158 kDa observed. 158.4 kDa calculated.
Physical form
Format: Purified
Other Notes
Concentration: Please refer to lot specific datasheet.
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- UPC:
- 12352200
- Condition:
- New
- Availability:
- 3-5 Days
- Weight:
- 1.00 Ounces
- HazmatClass:
- No
- MPN:
- MABS1291
- Temperature Control Device:
- Yes
