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SAB4200756

Sigma-Aldrich

Anti-AsCas12a (Cpf1) antibody, Mouse monoclonal

clone AsCpf-11, purified from hybridoma cell culture

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

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

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This Item
SAB4200777SAB4200751SAB4200701
clone

AsCpf-11, monoclonal

clone

LbCpf1, monoclonal

clone

10C11-A12, monoclonal

clone

7A9-3A3, monoclonal

antibody form

purified from hybridoma cell culture

antibody form

purified from hybridoma cell culture

antibody form

purified from hybridoma cell culture

antibody form

purified from hybridoma cell culture

technique(s)

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

technique(s)

immunoblotting: 0.6-1.2 μg/mL using human HEK-293T cells over-expressing LbCpf1 protein, immunofluorescence: 0.25-0.5 μg/mL using human HEK-293T cells over-expressing LbCpf1 protein, immunoprecipitation (IP): 1-2.5 μg/test using lysate of human HEK-293T cells over-expressing LbCpf1 protein

technique(s)

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

technique(s)

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

UniProt accession no.

U2UMQ6

UniProt accession no.

A0A182DWE3

UniProt accession no.

Q99ZW2

UniProt accession no.

-

Quality Level

200

Quality Level

200

Quality Level

200

Quality Level

200

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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.

Storage Class

10 - Combustible liquids

flash_point_f

Not applicable

flash_point_c

Not applicable


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Yuchen Liu et al.
Nature communications, 8(1), 2095-2095 (2017-12-14)
The catalytically dead Cpf1 endonuclease from Acidaminococcus sp. BV3L6 (dAsCpf1) has been used to construct effective transcriptional repressors in bacteria and plants. However, it is still unclear if dAsCpf1 can function in human cells as a transcriptional regulator or a
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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