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Merck

SAB4200701

Sigma-Aldrich

Anti-CRISPR/Cas9 antibody, Mouse monoclonal

clone 7A9-3A3, purified from hybridoma cell culture

別名:

CRISPR-associated protein-9 nuclease, Crisper, Crispr RNA

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About This Item

UNSPSCコード:
12352203
NACRES:
NA.41
クローン:
7A9-3A3, monoclonal
application:
IF
IP
テクニック:
immunoblotting: 1-2 μg/mL using whole extracts of human HEK-293T cells over-expressing CAS9 protein
immunofluorescence: 1-2 μg/mL using human HEK-293T cells over-expressing CAS9 protein.
immunoprecipitation (IP): 5-10 μg using whole extract of human HEK-293T cells over-expressing CAS9 protein
citations:
12

由来生物

mouse

品質水準

抗体製品の状態

purified from hybridoma cell culture

抗体製品タイプ

primary antibodies

クローン

7A9-3A3, monoclonal

フォーム

buffered aqueous solution

包装

antibody small pack of 25 μL

濃度

~1 mg/mL

テクニック

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

アイソタイプ

IgG1

輸送温度

dry ice

保管温度

−20°C

ターゲットの翻訳後修飾

unmodified

詳細

CAS9, also known as clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein-9 nuclease , cas5 and csn1, 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 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 targets genes by cleaving the foreign DNA in a sequence-dependent manner.2 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, reveling DSB sites in the genome and in ChIP (Chromatin Immunoprecipitation) related assays.

免疫原

a recombinant protein within the N-terminal region of Streptococcus pyogene Cas9

アプリケーション

Monoclonal Anti-CRISPR/CAS9 recognizes over-expression systems containing CAS9 construct. 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 does not cross react with FnCas9 from Francisella novicida bacteria and Cpf1 proteins from Acidaminococcus sp. (strain BV3L6) and Lachnospiraceae bacterium ND2006.

生物化学的/生理学的作用

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.

物理的形状

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

保管および安定性

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.

その他情報

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.

免責事項

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.

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保管分類コード

12 - Non Combustible Liquids

WGK

WGK 1

引火点(°F)

Not applicable

引火点(℃)

Not applicable


適用法令

試験研究用途を考慮した関連法令を主に挙げております。化学物質以外については、一部の情報のみ提供しています。 製品を安全かつ合法的に使用することは、使用者の義務です。最新情報により修正される場合があります。WEBの反映には時間を要することがあるため、適宜SDSをご参照ください。

Jan Code

SAB4200701-VAR:
SAB4200701-BULK:
SAB4200701-25UL:
SAB4200701-100UL:


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試験成績書(COA)

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文書ライブラリにアクセスする

L G V Fernandes et al.
Scientific reports, 9(1), 1839-1839 (2019-02-14)
Leptospirosis is a worldwide zoonosis caused by pathogenic bacteria of the genus Leptospira, which also includes free-living saprophyte strains. Many aspects of leptospiral basic biology and virulence mechanisms remain unexplored mainly due to the lack of effective genetic tools available
Yunjian Wu et al.
Molecular oncology, 17(3), 469-486 (2023-01-07)
Reciprocal interactions between prostate cancer cells and carcinoma-associated fibroblasts (CAFs) mediate cancer development and progression; however, our understanding of the signalling pathways mediating these cellular interactions remains incomplete. To address this, we defined secretome changes upon co-culture of prostate epithelial
Jana Ordon et al.
Plant methods, 19(1), 30-30 (2023-03-29)
In plant genome editing, RNA-guided nucleases such as Cas9 from Streptococcus pyogenes (SpCas9) predominantly induce small insertions or deletions at target sites. This can be used for inactivation of protein-coding genes by frame shift mutations. However, in some cases, it
Alicia Davis et al.
Molecular therapy. Methods & clinical development, 25, 158-169 (2022-04-12)
Hypoxia is a characteristic feature of solid tumors that contributes to tumor aggressiveness and is associated with resistance to cancer therapy. The hypoxia inducible factor-1 (HIF-1) transcription factor complex mediates hypoxia-specific gene expression by binding to hypoxia-responsive element (HRE) sequences
Yen-Ting Liu et al.
Molecular and cellular biology, 43(3), 115-129 (2023-03-22)
CDKN2A/B deletion or silencing is common across human cancer, reinforcing the general importance of bypassing its tumor suppression in cancer formation or progression. In rhabdomyosarcoma (RMS) and neuroblastoma, two common childhood cancers, the three CDKN2A/B transcripts are independently expressed to

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