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SAB4200777

Sigma-Aldrich

Anti-LbCas12a (Cpf1) antibody, Mouse monoclonal

clone LbCpf1, purified from hybridoma cell culture

Synonym(s):

Anti-CRISPR-associated endonuclease AsCas12a from Lachnospiraceae bacterium ND2006, Anti-LbCpf1 antibody, Mouse monoclonal

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About This Item

UNSPSC Code:
12352203
NACRES:
NA.41

biological source

mouse

Quality Level

antibody form

purified from hybridoma cell culture

antibody product type

primary antibodies

clone

LbCpf1, monoclonal

form

buffered aqueous solution

mol wt

~135 kDa

concentration

~1.0 mg/mL

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

isotype

IgG1

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. Depending on the architecture of the effector-CRISPR RNA (crRNA) interference module, different CRISPR-Cas systems could be assigned into two classes1: 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. 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 cleavage site.
LbCpf1, Cpf1 from Lachnospiraceae bacterium ND2006, was examined together with 15 members of Cpf1 nuclease family and proved to mediate efficient genome editing in HEK293FT cells with improved results compared to SpCas9.5 According to thr crystal structure, LbCpf1 has a triangle-shaped architecture with a large positively charged channel at the centre.7 The crRNA binding was shown to induce the pronounced structural rearrangements of LbCpf1, leading to formation of a substrate-binding conformation of LbCpf1. Over-expressed in plant cells, LbCpf1 demonstrated more than 10-fold transcriptional repression of the target gene.
Monoclonal anti-LbCpf1 antibody can provide a useful tool for genome editing research, such as detecting and monitoring LbCpf1 positively transfected cells.

Immunogen

Recombinant LbCpf1

Application

Monoclonal Anti-LbCpf1 specifically recognizes Cpf1 from Lachnospiraceae bacterium ND2006. The product may be used in several immunochemical techniques including Immunoblotting (∼135 kDa), Immunofluorescence and Immunoprecipitation. Monoclonal Anti-LbCpf1 does not cross react with Cpf1 from Acidaminococcus sp. (strain BV3L6) ), SpCas9 from Streptococcus pyogenes bacteria and FnCas9 from Francisella novicida bacteria.

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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WGK

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Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Regulatory Information

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Certificates of Analysis (COA)

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A modified Agrobacterium-mediated transformation for two oomycete pathogens.
Wang, et al.
PLoS Pathogens, 19, e1011346-e1011346 (2023)
Zuriñe Antón et al.
Journal of cell science, 133(18) (2020-08-28)
Current methodologies for targeting the mitochondrial genome for research and/or therapy development in mitochondrial diseases are restricted by practical limitations and technical inflexibility. A molecular toolbox for CRISPR-mediated mitochondrial genome editing is desirable, as this could enable targeting of mtDNA
Michael Aregger et al.
Nature protocols, 16(10), 4722-4765 (2021-09-12)
CRISPR-based forward genetic screening represents a powerful approach for the systematic characterization of gene function. Recent efforts have been directed toward establishing CRISPR-based tools for the programmable delivery of combinatorial genetic perturbations, most of which are mediated by a single

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