Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| M | WB, ICC | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MAB131872 |
| Description | Anti-Cas9 Antibody, clone 6F7 |
| Alternate Names |
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| Background Information | CRISPR-associated endonuclease Cas9 (UniProt: J7RUA5; also known as SaCas9, Cas9) is encoded by the Cas9 gene in Streptococcus aureus. Cas9 belongs to subtype II-A subfamily with one HNH Cas9-type domain. CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids). CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. Cas9 has a bilobed architecture with a recognition lobe (residues 41-425) and a discontinuous nuclease lobe (NUC, residues 1-40 and 453-1053); the crRNA-target DNA lies in a channel between the 2 lobes. The NUC lobe has 2 endonuclease domains. The discontinuous RuvC-like domain in NUC cleaves the target DNA non-complementary to crRNA while the HNH nuclease domain cleaves the target DNA complementary to crRNA. Cas9 serves as a RNA-guided DNA endonuclease that cleaves double-stranded DNA targets with a protospacer adjacent motif (PAM) and complementarity to the guide RNA. The homologous region of the Streptococcus aureus is not closely related in amino acid sequence in S. aureus, hence this antibody does not recognize that protein. |
| Product Information | |
|---|---|
| Format | Purified |
| Presentation | Purified mouse monoclonal antibody IgG1 in PBS with 0.03% sodium azide and 50% glycerol. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-Cas9, clone 6F7, Cat. No. MAB131872, is a highly specific mouse monoclonal antibody that targets CRISPR-associated endonuclease Cas9 and has been tested in Immunocytochemistry and Western Blotting. |
| Key Applications |
|
| Application Notes | Immunocytochemistry Analysis: A 1:1,000 dilution from a representative lot detected Cas9 in HEK293 cells which overexpress fusion protein containing GFP and C-terminus of Cas9 from S. aureus (Courtesy of Dr Gerry Shaw). Western Blotting Analysis: A representative lot detected Cas9 in HEK293 overexpressing fusion protein containing GFP and C-terminus of Cas9 from S. aureus (Courtesy of Dr Gerry Shaw). |
| Biological Information | |
|---|---|
| Immunogen | A recombinant fragement corresponding to 251 amino acids from the C-terminal region of Staphylococcus aureus. Expressed in and purified from E. coli. |
| Epitope | C-terminus |
| Clone | 6F7 |
| Concentration | Please refer to lot specific datasheet. |
| Host | Mouse |
| Specificity | Clone 6F7 specifically detects Cas9 in Staphylococcus aureus. It does not recognize Cas9 in Streptococcus pyogenes. |
| Isotype | IgG1κ |
| Species Reactivity |
|
| Species Reactivity Note | Staphylococcus aureus. |
| Antibody Type | Monoclonal Antibody |
| Gene Symbol |
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| Purification Method | Affinity Purified |
| UniProt Number |
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| Molecular Weight | ~53 kDa observed (truncated form). Uncharacterized bands may be observed in some lysate(s). |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blotting in HEK293 cell lysates overexpressing a fusion protein containing GFP and C-terminus of Cas9 from Staphylococcus aureus. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Cas9 in 10 µg of HEK293 cell lysates overexpressing a fusion protein containing GFP and c-terminus of Cas9 from Staphylococcus aureus. |
| Usage Statement |
|
| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. |
| Packaging Information | |
|---|---|
| Material Size | 100 μg |