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SAB4200756

Sigma-Aldrich

Anti-AsCas12a (Cpf1) antibody, Mouse monoclonal

clone AsCpf-11, purified from hybridoma cell culture

Synonym(s):

Anti-AsCpf1 antibody, Mouse monoclonal, Anti-CRISPR-associated endonuclease AsCas12a from Acidaminococcus sp. (strain BV3L6)

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About This Item

UNSPSC Code:
12352203
NACRES:
NA.43

biological source

mouse

Quality Level

antibody form

purified from hybridoma cell culture

antibody product type

primary antibodies

clone

AsCpf-11, monoclonal

form

buffered aqueous solution

mol wt

~135 kDa

concentration

~1.0 mg/mL

technique(s)

immunoblotting: 1.25-2.5 μg/mL using purified recombinant AsCpf1 produced in E. coli
immunofluorescence: 1.25-2.5 μg/mL using human HEK-293T cells over-expressing AsCpf1 protein
immunoprecipitation (IP): 2.5-5 μg/test using lysate of human HEK-293T cells over-expressing AsCpf1 protein

isotype

IgG2a

UniProt accession no.

shipped in

dry ice

storage temp.

−20°C

target post-translational modification

unmodified

General description

Clustered, regularly interspaced, short palindromic repeat (CRISPR) systems, encode RNA-guided endonucleases that are essential for bacterial adaptive immunity. Ddepending on the architecture of the effector-CRISPR RNA (crRNA) interference module, the ddifferent CRISPR-Cas systems could be assigned into two classes: Class-1 systems of multi-subunit complex such as Cascade and Class-2 systems of single enzyme, such as Cas9.
Cpf1 (CRISPR from Prevotella and Francisella 1) belongs to Class-2 type V CRISPR-Cas endonuclease system.4-5 Cpf1 comprise several differences from Cas9 protein including cleavage with 5′overhangs, a shorter guide RNA and a longer distance between the seed sequence and the cleavage site.
AsCpf1, Cpf1 from Acidaminococcus sp. (strain BV3L6), was examined together with 15 members of Cpf1 nuclease family and shown to mediate efficient genome editing in HEK293FT cells with an improved results compared to SpCas9.5 AsCpf1 crystal structure in a complex with crRNA and partially duplexed target DNA, shows that AsCpf1 acts as a monomer thus identifying a unique mechanism employed by AsCpf1 for target recognition.

Immunogen

recombinant Cpf1 from Acidaminococcus sp. (strain BV3L6)

Application

Monoclonal Anti-AsCpf1 recognizes Cpf1 from Acidaminococcus sp. (strain BV3L6). The product has been used in several immunochemical techniques including
  • immunoblotting (~135 kDa),
  • immunofluorescence
  • immunoprecipitation

Monoclonal anti-AsCpf1 antibody can provide a useful tool for genome editing research such as detecting and monitoring AsCpf1 positively transfected cells.

Physical form

Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide.

Storage and Stability

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.

Other Notes

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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Storage Class Code

10 - Combustible liquids

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Pu Gao et al.
Cell research, 26(8), 901-913 (2016-07-23)
CRISPR-Cas9 and CRISPR-Cpf1 systems have been successfully harnessed for genome editing. In the CRISPR-Cas9 system, the preordered A-form RNA seed sequence and preformed protein PAM-interacting cleft are essential for Cas9 to form a DNA recognition-competent structure. Whether the CRISPR-Cpf1 system
Giedrius Gasiunas et al.
Proceedings of the National Academy of Sciences of the United States of America, 109(39), E2579-E2586 (2012-09-06)
Clustered, regularly interspaced, short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems provide adaptive immunity against viruses and plasmids in bacteria and archaea. The silencing of invading nucleic acids is executed by ribonucleoprotein complexes preloaded with small, interfering CRISPR RNAs (crRNAs) that act
Hui K Kim et al.
Nature methods, 14(2), 153-159 (2016-12-20)
CRISPR from Prevotella and Francisella 1 (Cpf1) is an effector endonuclease of the class 2 CRISPR-Cas (clustered regularly interspaced short palindromic repeats-CRISPR-associated proteins) gene editing system. We developed a method for evaluating Cpf1 activity, based on target sequence composition in
Eva Schunder et al.
International journal of medical microbiology : IJMM, 303(2), 51-60 (2013-01-22)
Francisella tularensis is a zoonotic agent and the subspecies novicida is proposed to be a water-associated bacterium. The intracellular pathogen F. tularensis causes tularemia in humans and is known for its potential to be used as a biological threat. We
Engineering cell signaling using tunable CRISPR-Cpf1-based transcription factors
Liu Y, et al.
Nature Communications, 8(1), 2095-2095 (2017)

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