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MAB131872

Sigma-Aldrich

Anti-Cas9 Antibody, clone 6F7

clone 6F7, from mouse

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Synonym(s):
CRISPR-associated endonuclease Cas9, SaCas9
eCl@ss:
32160702

biological source

mouse

Quality Level

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

6F7, monoclonal

species reactivity

Staphylococcus aureus, mouse

technique(s)

immunocytochemistry: suitable
western blot: suitable

isotype

IgG1κ

UniProt accession no.

shipped in

ambient

target post-translational modification

unmodified

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MAC142MAC141MABE1668
Anti-Cas9 Antibody, clone 6F7 clone 6F7, from mouse

MAB131872

Anti-Cas9 Antibody, clone 6F7

clone

6F7, monoclonal

clone

6H4, monoclonal

clone

5F1, monoclonal

clone

6G12-H11, monoclonal

biological source

mouse

biological source

mouse

biological source

mouse

biological source

mouse

species reactivity

Staphylococcus aureus, mouse

species reactivity

Staphylococcus aureus, bacteria

species reactivity

Staphylococcus aureus, bacteria

species reactivity

Streptococcus pyogenes

UniProt accession no.

J7RUA5

UniProt accession no.

J7RUA5

UniProt accession no.

J7RUA5

UniProt accession no.

Q99ZW2

Quality Level

100

Quality Level

100

Quality Level

100

Quality Level

100

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General description

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.

Specificity

Clone 6F7 specifically detects Cas9 in Staphylococcus aureus. It does not recognize Cas9 in Streptococcus pyogenes.

Immunogen

A recombinant fragement corresponding to 251 amino acids from the C-terminal region of Staphylococcus aureus. Expressed in and purified from E. coli.

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.
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).

Quality

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.

Target description

~53 kDa observed (truncated form). Uncharacterized bands may be observed in some lysate(s).

Physical form

Format: Purified
Purified mouse monoclonal antibody IgG1 in PBS with 0.03% sodium azide and 50% glycerol.

Other Notes

Concentration: Please refer to lot specific datasheet.

Storage Class

10 - Combustible liquids

wgk_germany

WGK 2


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Pin Lyu et al.
Nucleic acids research, 47(17), e99-e99 (2019-07-13)
Transient expression of the CRISPR/Cas9 machinery will not only reduce risks of mutagenesis from off-target activities, but also decrease possible immune response to Cas9 protein. Building on our recent developing of a system able to package up to 100 copies
Xingang Yao et al.
Journal of extracellular vesicles, 10(5), e12076-e12076 (2021-03-23)
Transient delivery of CRISPR-based genome editing effectors is important to reduce off-target effects and immune responses. Recently extracellular vesicles (EVs) have been explored for Cas9 ribonucleoprotein (RNP) delivery. However, lack of mechanisms to enrich RNPs into EVs limited the efficiency
Baisong Lu et al.
Nucleic acids research, 47(8), e44-e44 (2019-02-14)
The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system discovered using bacteria has been repurposed for genome editing in human cells. Transient expression of the editor proteins (e.g. Cas9 protein) is desirable to reduce the risk of mutagenesis from

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