| Field | Specification |
|---|---|
| Product Type | |
| Shipping | |
| Source | Recombinant (E. coli) |
| Storage |
The AapCas12b nuclease (also known as C2c1) is a DNA endonuclease mediated by tracrRNA:crRNA (or sgRNA), derived from the thermophilic bacterium Alicyclobacillus acidophilus. In the presence of a PAM (TTN) sequence at the target double-stranded DNA (dsDNA), it specifically cleaves the target dsDNA, causing double-strand breaks and generating sticky ends. AapCas12b can specifically cleave single-stranded DNA targets independent of the PAM sequence. Both double-stranded and single-stranded DNA targets can activate the trans-cleavage activity of AapCas12b. When the AapCas12b enzyme forms a ternary complex with sgRNA and target DNA, it becomes activated for nonspecific ssDNA trans-cleavage activity, cutting any arbitrary sequence of ssDNA in the system. The optimal cleavage reaction temperature for AapCas12b is 60oC, which makes it more heat-resistant than AacCas12b, and more suitable for use with LAMP to develop isothermal amplification/CRISPR-Cas detection systems.
Features
Broad reaction temperature range: Exhibits cleavage activity within the temperature range of 37~65oC
Low nuclease residue: No residual exonuclease, nickase, or RNase
Cis-cleavage activity: Highly Effective Cleavage of Double-Stranded DNA in Vitro
Trans-cleavage activity: High trans-cleavage activity, suitable for nucleic acid detection
Applications
CRISPR/Cas gene editing
Diagnostic and detection based on the CRISPR/Cas system
Other detection applications combined with isothermal nucleic acid amplification technologies (RPA and LAMP), etc
Specifications
|
Molecular weight |
130 KDa |
|
Concentration |
10 µM |
|
Purity |
>95% (SDS-PAGE) |
|
Unit Definition |
The quantity of Cas12b enzyme required to cleave 1 pmol of ssDNA probe within 1 minute at 60oC in a total reaction volume of 20 μL, containing 1×reaction buffer, is defined as 1 U |
Glycerol Content |
Contains Glycerol |
Components
|
Components No. |
Name |
14808ES65 |
14808ES80 |
|
14808-A |
AapCas12b Nuclease (10 μM) |
100 μL |
|
|
14808-B |
10×Reaction buffer |
1 mL |
1 mL |
Shipping and Storage
This product should be stored at -25 ~ -15oC for 2 years.
Figures
Figure 1. Cis-cleavage Activity Verification of AapCas12b Nuclease
Note: In a 20 μL reaction system containing Target dsDNA with a PAM sequence, sgRNA, Cas12b, and 1×reaction buffer, the mixture was incubated at 60oC for 30 minutes and then inactivated at 85oC for 5 minutes. The agarose gel electrophoresis results showed that three batches of AapCas12b Nuclease were all capable of effectively cleaving the dsDNA, with cleavage efficiency comparable to that of the imported brand T*.
Figure 2. AapCas12b Nuclease Trans-cleavage Activity Test Results
Note: In a 20 μL reaction system containing Target dsDNA with a PAM sequence, sgRNA, Cas12b, Report ssDNA fluorescent probe, and 1×reaction buffer, the mixture was incubated at 60oC for 1 hour, and the fluorescence signal was collected every 30 seconds. The results showed that in the presence of Target DNA, sgRNA, and Cas12b together, the Report ssDNA fluorescent probe could be cleaved, thereby releasing the fluorescence signal.
Store this enzyme at -20°C and avoid repeated freeze-thaw cycles to preserve catalytic activity. The product is shipped Dry Ice and remains stable for up to one year from the date of manufacture when stored under recommended conditions. Aliquoting the stock solution into single-use volumes is recommended for enzymes used infrequently to minimize thermal cycling of the bulk stock.
CRISPR/Cas gene editing Diagnostic and detection based on the CRISPR/Cas system Other detection applications combined with isothermal nucleic acid amplification technologies (RPA and LAMP), etc. Always verify compatibility with your specific template, buffer, and downstream workflow.
Nuclease activity is expressed in µg/µL; cleavage efficiency is validated as the percentage of target DNA cut within 60 min at 37°C using a defined guide RNA and target plasmid substrate.
This enzyme is produced as Recombinant (E. coli) and supplied as a Research Use Only (RUO) reagent. Each lot is subjected to activity assay, purity assessment by SDS-PAGE, and functional validation prior to release. A Certificate of Analysis (CoA) and Safety Data Sheet (SDS) are available on request.
On-target efficiency is primarily determined by guide RNA (gRNA) design, target accessibility within chromatin, and PAM sequence context. Off-target cleavage is influenced by sequence complementarity mismatches (particularly in the seed region proximal to the PAM), enzyme concentration, and delivery method. High-fidelity Cas9 variants with engineered positional charge substitutions significantly reduce off-target activity (>99% reduction) while maintaining on-target cleavage efficiency comparable to wild-type enzyme.
Yeasen Biotechnology supports custom enzyme solutions across multiple service lines — from GMP-grade bulk supply to directed enzyme engineering. Contact BioHippo to discuss requirements and initiate a project inquiry.
▶ GMP-Grade & Bulk Supply
Select Yeasen enzymes are available in GMP grade, manufactured in an ISO 13485-certified UCF.ME™ ultra-clean molecular enzyme facility with FDA Drug Master File (DMF) support.
- GMP-grade release testing and CoA documentation
- ISO 13485-certified production facility
- Scalable from milligram to multi-gram quantities
- Consistent lot-to-lot activity specifications
▶ Glycerol-Free & Custom Formulation
Glycerol-free enzyme formats are available for applications requiring lyophilization compatibility, liquid handling automation, or direct IVD master mix integration.
- Glycerol-free liquid format (standard and custom buffers)
- Lyophilization-ready enzyme preparation
- Custom reaction buffer optimization for specific assay conditions
- Compatible with freeze-drying workflows for point-of-care formats
▶ Molecular IVD RDC Service
Yeasen's Research and Development Contracting (RDC) team delivers end-to-end solutions for molecular diagnostic product development, covering enzyme selection through clinical validation support.
- Enzyme selection and performance matching
- Primer/probe design and reaction buffer optimization
- Sensitivity, specificity, and precision validation studies
- Stability studies and SNP evaluation
- Instrument platform compatibility assessment
▶ ZymeEditor™ Enzyme Engineering
Yeasen's proprietary ZymeEditor™ directed evolution and rational design platform enables the development of custom enzyme variants with tailored performance characteristics not available in off-the-shelf products.
- Directed evolution for enhanced thermostability, processivity, or fidelity
- Rational design for altered substrate specificity or cofactor requirements
- Library screening from Yeasen's proprietary enzyme variant collection
- Scale-up to commercial quantities upon candidate confirmation
ⓘ Customization services are fulfilled by Yeasen Biotechnology. Lead times and minimum order quantities vary by service type. Contact BioHippo for project scoping and pricing.
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