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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| A | ICC, WB | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MABE1669 |
| Description | Anti-Cas9 Antibody, NT Antibody, clone 8C1-F10 |
| Alternate Names |
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| Background Information | 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 M1s 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. |
| Product Information | |
|---|---|
| Format | Purified |
| Presentation | Purified mouse IgG2bκ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-Cas9, NT, clone 8C1-F10 , Cat. No. MABE1669, is a highly specific mouse monoclonal antibody that targets CRISPR-associated endonuclease Cas9/Csn1 and has been tested in Immunocytochemistry and Western Blotting. |
| Key Applications |
|
| Application Notes | 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. |
| Biological Information | |
|---|---|
| Immunogen | His-tagged recombinant Cas9 N-terminal fragment excluding the active site RuvC-I. |
| Clone | 8C1-F10 |
| Concentration | Please refer to lot specific datasheet. |
| Host | Mouse |
| Specificity | Clone 8C1-F10 targets an epitope within Cas9 N-terminal region outside the active site RuvC-I. |
| Isotype | IgG2bκ |
| Species Reactivity |
|
| Species Reactivity Note | Streptococcus pyogenes. |
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Gene Symbol |
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| Purification Method | Protein G purified. |
| UniProt Number |
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| Molecular Weight | ~160 kDa observed. 158.4 kDa calculated. Uncharacterized bands may be observed in some lysate(s). |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blotting in NIH3T3 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. |
| Usage Statement |
|
| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at 2-8°C from date of receipt. |
| Packaging Information | |
|---|---|
| Material Size | 100 μg |