MAC148

Anti-Cas9 Antibody, clone 8G4

• Synonym(s): CRISPR-associated endonuclease Cas9/Csn1, SpCas9, SpyCas9
• UNSPSC Code: 12352203
• NACRES: NA.43

Properties

• biological source: rat
• Quality Level: 200
• antibody form: purified antibody
• antibody product type: primary antibodies
• clone: 8G4, monoclonal
• mol wt: calculated mol wt 158.44 kDa; observed mol wt ~210 kDa
• purified by: using protein G
• species reactivity: all
• packaging: antibody small pack of 100
• technique(s): immunofluorescence: suitable; western blot: suitable
• isotype: IgG2aλ
• epitope sequence: Unknown
• Protein ID accession no.: NP_269215
• UniProt accession no.: Q99ZW2
• storage temp.: 2-8°C
• Gene Information: Streptococcus pyogenes ... Cas9(69900935)

Description

Specificity

Clone 8G4 is a rat monoclonal antibody that specifically detects Cas9.

Immunogen

His-tagged, full-length, recombinant Cas9 from Streptococcus pyogenes.

Application

Quality Control Testing

Evaluated by Western Blotting in lysate from HeLa cells overexpressing Cas9.

Western Blotting Analysis: A 1:500 dilution of this antibody detected Cas9 in lysate from HeLa cells overexpressing Cas9.

Tested Applications

Immunofluorescence Analysis: A 1:500 dilution from a representative lot detected Cas9 in Cell line stably expressing Cas9-NLS (A) versus wild type parental cell line (B) (Courtesy of Germán Camargo Ortega (Institute of Stem Cell Research, Helmholtz Munich,. Germany).

Western Blotting Analysis: A representative lot detected Cas9 in Western Blotting application. (Camargo Ortega, G., et al. (2019). Nature. 567(7746):113-117).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Target description

CRISPR-associated endonuclease Cas9/Csn1 (UniProt: Q99ZW2; also known as SpCas9, SpyCas9, Cas9) is encoded by the Cas9 gene (Gene ID: 69900935) in Streptococcus pyogenes. 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 bi-lobed 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. (Ref.: Fonfara, I., et al. (2014). Nucleic Acids Res. 42(4); 2577-2590; Jinek, M., et al. (2012). Science. 337(6096); 816-821).

Physical form

Purified rat monoclonal antibody IgG2a in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Reconstitution

0.5 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommended storage: +2°C to +8°C.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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.

Safety Information

• Storage Class: 10 - Combustible liquids
• wgk_germany: WGK 1
• flash_point_f: Not applicable
• flash_point_c: Not applicable