Description
Recombinant Cas12a Protein for Precision Gene Detection Technologies: LbCas12a (LbCpf1)
- Recombinant LbCas12a (LbCpf1) for forming RNP complexes with guide-RNA
- Highly stable and free of RNAses
- Highly purified and extremely active (staggered double strand DNA cleavage)
- Trans ssDNA cleavage activity can be used to cleave ssDNA reporters generating detectable signals
- Bulk Pricing available – please contact us for a quote
SignalChem’s highly purified recombinant Lachnospiraceae bacterium Cas12a (LbCas12a, LbCpf1) protein (>90% purity) without nuclear localization signal (NLS) is used for CRISPR-based gene detection technologies and for CRISPR-mediated in vitro applications. The enzyme was expressed in E. coli and purified with a his tag.
LbCas12a protein combines with guide RNA (gRNA) to form a stable ribonucleoprotein (RNP) complex that enables the inactive nuclease to be guided to the specific double stranded DNA (dsDNA) site. Once guided to the specific cleavage site, LbCas12a becomes activated to specifically and precisely cleave the dsDNA and to indiscriminately trans-cleave single stranded DNA (ssDNA) nearby. A short single-stranded DNA labeled with a fluorophore and a quencher on the two termini can be used as a reporter to generate a detectable fluorescence signal for highly precise detection of the guide RNA-targeted dsDNA sequence. The highly precise detection system can be combined with pre-amplification procedures including isothermal amplifications such as RPA (Recombinase Polymerase Amplification) and LAMP, to achieve one-pot reaction systems, suitable for field-testing, and offering many advantages over qPCR-based detection.
Moreover, Cas12a has intrinsic RNase activity that allows processing of its own crRNA array enabling multiplex editing for combinatorial genetic screening.
Compared to AsCas12a, which is another Cas12a ortholog, LbCas12a has been shown to be more active at a lower range of temperatures from 22°C to 37°C in vitro and in cells, making it ideal for plants.
SPECIFICATIONS
PURITY | >90% (SDS-PAGE gel analysis)
ENZYME SOURCE | Lachnospiraceae sp. Cas12a is expressed in E. coli without nuclear localization signal (NLS), and is purified using a his tag.
QUANTITY | 100 ug, plus 1ml 10x Cas12 Reaction Buffer
STORAGE/SHIPPING CONCENTRATION | 0.1 ug/ul
STORAGE BUFFER | 50mM sodium phosphate, pH 7.5, 300mM NaCl, 150mM imidazole, 1mM DTT and 10% glycerol
RECOMMENDED STORAGE CONDITIONS | -80°C
EXPIRATION | 12 months from receipt when stored as directed. Avoid repeated freeze/thaw cycles.