AsCpf1/Cas12a (Strain BV3L6) (E1U7C) Rabbit mAb #19984

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Supporting Data

REACTIVITY
SENSITIVITY	Transfected Only
MW (kDa)	151
Source/Isotype	Rabbit IgG

Application Key:

WB-Western Blotting
IF-Immunofluorescence
F-Flow Cytometry

Product Information

Product Usage Information

Application	Dilution
Western Blotting	1:1000
Immunofluorescence (Immunocytochemistry)	1:400
Flow Cytometry (Fixed/Permeabilized)	1:400

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #33209.

Protocol

Specificity / Sensitivity

AsCpf1/Cas12a (Strain BV3L6) (E1U7C) Rabbit mAb recognizes transfected levels of total AsCpf1/Cas12a (Strain BV3L6) protein. This antibody does not cross-react with Cas9 (S. pyogenes), Cas9 (S. aureus), and FnCpf1/Cas12a (Strain U112) proteins.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu822 of Acidaminococcus sp. Cpf1/Cas12a (Strain BV3L6) protein.

Background

CRISPR-Cas (clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins) are RNA-guided nuclease effectors that are utilized for precise genome editing in mammalian systems (1). Cpf1/Cas12a (CRISPR from Prevotella and Francisella) proteins are members of the Class 2 CRISPR system (2). Class 2 CRISPR systems, such as the well characterized Cas9, rely on single-component effector proteins to mediate DNA interference (3). Cpf1/Cas12a endonucleases, compared to Cas9 systems, have several unique features that increase the utility of CRISPR-based genome editing techniques: 1) Cpf1/Cas12a-mediated cleavage relies on a single and short CRISPR RNA (crRNA) without the requirement of a trans-activating crRNA (tracrRNA), 2) Cpf1/Cas12a utilizes T-Rich protospacer-adjacent motif (PAM) sequences rather than a G-Rich PAM, and 3) Cpf1/Cas12a generates a staggered, rather than a blunt-ended, DNA double-stranded break (2). These features broaden the utility of using CRISPR-Cas systems for specific gene regulation and therapeutic applications. Several Cpf1/Cas12a bacterial orthologs have been characterized for CRISPR-mediated mammalian genome editing (2,4). AsCpf1 (Strain BV3L6)/Cas12a is a Cpf1/Cas12a enzyme derived from Acidaminococcus sp. BV3L6 (5,6).

Database Links

UniProt ID: U2UMQ6

Entrez-Gene Id:

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