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FastScan^™ LbCpf1 (Strain ND2006)/Cas12a ELISA Kit

FastScan^™ LbCpf1 (Strain ND2006)/Cas12a ELISA Kit #88197

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Supporting Data

REACTIVITY

All

Application Key:

ELISA-ELISA

Species Cross-Reactivity Key:

All-All Species Expected

Product Information

Product Description

The FastScan™ LbCpf1 (Strain ND2006)/Cas12a ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of LbCpf1 (Strain ND2006)/Cas12a protein. To perform the assay, sample is incubated with a capture antibody conjugated with a proprietary tag and a second detection antibody linked to HRP, forming a sandwich with LbCpf1 (Strain ND2006)/Cas12a protein in solution. This entire complex is immobilized to the plate via an anti-tag antibody. The wells are then washed to remove unbound material. TMB is then added. The magnitude of observed signal is proportional to the quantity of LbCpf1 (Strain ND2006)/Cas12a protein.

*Antibodies in this kit are custom formulations specific to kit.

IMPORTANT: This FastScan™ ELISA Kit requires 4 washes at Step 6 of the protocol.

Protocol

Specificity / Sensitivity

The FastScan™ LbCpf1 (Strain ND2006)/Cas12a ELISA Kit detects endogenous levels of LbCpf1 (Strain ND2006)/Cas12a protein, as shown in Figure 1. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Species Reactivity:

All Species Expected

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).~LbCpf1 (Strain ND2006)/Cas12a is a Cpf1/Cas12a enzyme derived from Lachnospiraceae bacterium ND2006 (2,4).

Database Links

UniProt ID: A0A182DWE3

Entrez-Gene Id:

Limited Uses

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For Research Use Only. Not For Use In Diagnostic Procedures.

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FastScan™ ELISA is a registered trademark of Cell Signaling Technology, Inc.

U.S. Patents 9,086,407, 9,261,500, and 9,476,874, foreign equivalents, and child patents deriving therefrom.

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FastScan™ LbCpf1 (Strain ND2006)/Cas12a ELISA Kit #88197