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Merck

SAB4200751

Sigma-Aldrich

Anti-CRISPR/Cas9 (C-terminal) antibody, Mouse monoclonal

clone 10C11-A12, purified from hybridoma cell culture

Sinónimos:

Anti-CRISPR-associated protein-9 nuclease, Anti-Crispr, Anti-Crsipr RNA

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About This Item

Código UNSPSC:
12352203
NACRES:
NA.41

forma del anticuerpo

purified from hybridoma cell culture

Nivel de calidad

tipo de anticuerpo

primary antibodies

clon

10C11-A12, monoclonal

Formulario

buffered aqueous solution

concentración

~1.00 mg/mL

técnicas

immunoblotting: 1-2 μg/mL using whole extracts of human HEK-293T cells over-expressing CAS9 protein
immunofluorescence: 1.25-2.5 μg/mL using human HEK-293T cells over-expressing CAS9 protein
immunoprecipitation (IP): 5-10 μg/test using whole extract of human HEK-293T cells over-expressing CAS9 protein

Nº de acceso UniProt

Condiciones de envío

dry ice

temp. de almacenamiento

−20°C

modificación del objetivo postraduccional

unmodified

Descripción general

CAS9, also known as clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein-9 nuclease, is the signature gene of the type II CRISPR -RuvC (RNase H-like fold) Cas system. CAS9 contains 1388 amino acids. This protein is predicted to contain a RuvC/ ribonuclease (RNase) H domain involved in CRISP RNA (crRNA) maturation and McrA/HNH signature domain involved in the DNA degradation step.
CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) belongs to the type II CRISPR/CAS9 system. It is part of an adaptive immune system of the Streptococcus pyogenes SF370, protecting from pathogens′ target genes by cleaving the foreign DNA in a sequence-dependent manner. The type II CRISPR/Cas system which has been adapted to expression in eukaryotic cells, consists of four genes including the Cas9 (CRISPR-associated proteins) nuclease, two noncoding CRISPR RNAs (crRNAs, or gRNA), trans-activating crRNA (tracrRNA) and a precursor crRNA (pre-crRNA) array. The pre-crRNA contains nuclease guide sequences (spacers) interspaced by identical direct repeats (DRs).
The Cas9 endonuclease can be engineered with a single gRNA, directing a DNA double-strand break (DSB) at a desired genomic location. Similar to DSBs induced by zinc finger nucleases (ZFNs), the cell then activates endogenous DNA repair processes, either non-homologous end joining (NHEJ) or homology-directed repair (HDR), to heal the targeted DSB.
In comparison to other genome-editing technologies such as designer zinc fingers (ZFs), transcription activator–like effectors (TALEs) and homing meganucleases, the CRISPR/CAS9 system is a scalable, affordable and easy to engineer. Therefore, the anti-CRISPR/CAS9 antibody can be a useful tool for detecting CRISPR/CAS9 positively transfected cells, revealing DSB sites in the genome and in ChIP (Chromatin Immunoprecipitation) related assays

Inmunógeno

Recombinant protein within the C-terminal region of Streptococcus pyogenes Cas9

Aplicación

Monoclonal Anti-CRISPR/CAS9 (C-terminal) recognizes CAS9 protein in CAS9 construct over-expression systems. The antibody successfully recognizes mutant Cas9 variants, including nickase Cas9 and dead Cas9 (dCas9). The antibody may be used in various immunochemical techniques including Immunoblotting, Immunofluorescence and Immunoprecipitation. Monoclonal Anti- CRISPR/CAS9 (C-terminal) does not cross react with FnCas9 from Francisella novicida bacteria and Cpf1 proteins from Acidaminococcus sp. (strain BV3L6) and Lachnospiraceae bacterium ND2006.

Acciones bioquímicas o fisiológicas

CAS9 plays a vital role in plasmid DNA interference. It is the only Cas protein needed to deliver resistance against foreign DNA. CAS9 stimulates both RNA-guided genome editing and gene regulation in various organisms, but it can facilitate only one activity at a time within any given cell.

Forma física

Supplied as a solution in 0.01 M phosphate buffered saline pH 7.4, containing 15 mM sodium azide as a preservative.

Almacenamiento y estabilidad

For continuous use, store at 2-8 °C for up to one month. For extended storage, freeze in working aliquots. Repeated freezing and thawing is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilution samples should be discarded if not used within 12 hours.

Otras notas

This product is for R&D use only, not for drug, household, or other uses.
In order to obtain best results in different techniques and preparations we recommend determining optimal working concentration by titration test.

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Código de clase de almacenamiento

10 - Combustible liquids

Punto de inflamabilidad (°F)

Not applicable

Punto de inflamabilidad (°C)

Not applicable


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Visite la Librería de documentos

Snigdha Poddar et al.
Frontiers in plant science, 13, 1084700-1084700 (2023-01-28)
The advancement of precision engineering for crop trait improvement is important in the face of rapid population growth, climate change, and disease. To this end, targeted double-stranded break technology using RNA-guided Cas9 has been adopted widely for genome editing in
Benjamin L Oakes et al.
Cell, 176(1-2), 254-267 (2019-01-12)
The ability to engineer natural proteins is pivotal to a future, pragmatic biology. CRISPR proteins have revolutionized genome modification, yet the CRISPR-Cas9 scaffold is not ideal for fusions or activation by cellular triggers. Here, we show that a topological rearrangement of Cas9
The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli
Sapranauskas R, et al.
Nucleic Acids Research, 39(21), 9275-9282 (2011)
Orthogonal Cas9 proteins for RNA-guided gene regulation and editing
Esvelt K M, et al.
Nature Methods, 10(11), 1116-1116 (2013)
Sybille Böhm et al.
Science advances, 6(34), eaba5614-eaba5614 (2020-09-03)
Catalytically inactive dCas9 fused to transcriptional activators (dCas9-VPR) enables activation of silent genes. Many disease genes have counterparts, which serve similar functions but are expressed in distinct cell types. One attractive option to compensate for the missing function of a

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