Cas13a, His-Tag (L. wadei) Recombinant

Only %1 left

Catalog #

101630

As low as $345 *

Cat # Size Price*

Qty

Bulk/Custom

Add to Favorites

*US Pricing only. For international pricing, please contact your local distributor.

●

Purchase

●

Description

Recombinant L. wadei LwCas13a (type VI-A CRISPR-associated RNA-guided ribonuclease Cas13a), encompassing amino acids 1-1152. This construct contains a C-terminal His Tag. Cas13a is an RNA-guided endonuclease that belongs to the class 2 type VI CRISPR-Cas system. When activated, Cas13a induces collateral cleavage of nearby non-targeted RNAs in a nonspecific manner.

●

Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

LwCas13a Collateral Cleavage Activity

Product Info

Storage and Usage

Citations

Species

Leptotrichia wadei

Construct

Cas13a (1-1152-His) (L. wadei)

Host Species/Expression System

E. coli

Purity

≥90%

Format

Aqueous buffer solution.

Formulation

50 mM Tris, pH 7.5, 600 mM NaCl, 2 mM DTT, and 10% glycerol

125 kDa

Genbank #

WP_021746774

Tag(s)

C-terminal His-tag

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

Varying amounts of LwCas13a activity was measured using a CRISPR-based fluorescent reporter assay for optimal results. Target RNA cutting and collateral RNase activity was activated using RNA-guided RNA Binding to Cas13a. Emission of fluorescent signal is due to the degradation of the reporter substrate upon cleavage.

Active Cas13 was thawed on ice while 1X Reaction Buffer containing 20 mM HEPES, pH 7.0, 50 mM KCl, 5 mM MgCl₂, and 0.1 mg/ml BSA, guide RNA (target-specific spacer sequence), target RNA activator (complementary sequence to crRNA) were equilibrated to room temperature. Next three working solutions of Active Cas13 (4X final concentration) guide RNA (4X final concentration) and activator/reporter mix containing RNA activator and reporter substrate (2X final concentration), were prepared using 1X Reaction Buffer. 10 µl of 4X active Cas12 and 10 µl of 4x guide RNA were then preincubated in half the area of a solid black 96-well plate for 5 minutes at room temperature. After preincubation, 20 µl of 2X activator/reporter mix was added to plate and placed on shaking incubator for 1 min. The plate was then sealed and incubated at 37°C for 10-30 minutes. Plate was then equilibrated to room temperature, plate sealer removed and fluorescence read on a microplate reader. Negative control was measured by replacing enzyme working solution with equal volume of assay buffer.

Instructions for Use

Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage.

Applications

Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Shipping Temperature

-80°C

Warnings

Avoid multiple freeze/thaw cycles.

Scientific Category

CRISPR

Regulatory Status

Research Use Only

Skip to the end of the images gallery

Skip to the beginning of the images gallery

●

Datasheets/Protocols

101630 lot 230110

Datasheets provide example lot-specific information. Contact us to request information on currently available lots.

● General Protocols

● FAQs

● Contact Technical Service

●

Cas13a, His-Tag (L. wadei) Recombinant

PRODUCTS

SERVICES

RESOURCES

COMPANY