| Field | Specification |
|---|---|
| Mfr No | |
| Product Type | |
| Shipping | |
| Source | Recombinant (E. coli) |
| Storage |
Product description
Alkaline Phosphatase, can remove the 5' phosphate groups from DNA and RNA. It is commonly used to prevent the self-ligation of vectors. In molecular cloning experiments, DNA ligase requires the presence of phosphate groups to catalyze DNA ligation. After digestion with restriction enzymes, vectors retain a phosphate group at the cleavage site. However, after dephosphorylation by Alkaline Phosphatase, the 5' end of the vector lacks a phosphate group and thus cannot connect with its own 3' end. As a result, the self-ligation of the vector, which would otherwise preferentially occur in the ligation reaction, is prevented. This increases the insertion rate of the target fragment. Additionally, Alkaline Phosphatase can be used to prepare DNA templates for 5' end labeling.
This product is a shrimp-derived Alkaline Phosphatase that can degrade almost all phosphate monoesters. However, it cannot hydrolyze phosphate diesters or triesters. The enzyme acts on the terminal dephosphorylation, regardless of whether the termini are sticky or blunt. SAP can also be used to degrade dNTPs in PCR reactions, which is useful for subsequent sequencing and SNP analysis.
Features
- Removal of phosphate groups from DNA and RNA ends. Preventing self-ligation of vectors.
- Preparation of DNA templates for 5' end labeling.
- Degradation of dNTPs in PCR reactions.
Applications
- Molecular cloning
- DNA sequencing
- SNP analysis
Components
|
Components No. |
Name |
10322ES72 |
|
10322-A |
Shrimp Alkaline Phosphatase (SAP) (1 U/µL) |
250 U |
|
10322-B |
10×SAP buffer |
1 mL |
Specifications
|
From Pichia pastoris, the shrimp alkaline phosphatase gene was cloned from Pandalus borealis. |
|
|
Quality Control |
Free of ribonuclease and deoxyribonuclease contamination. |
|
Purity |
＞95% |
|
Thermal Inactivation |
Incubation at 65℃ for 5 min results in complete and irreversible loss of activity. |
|
Unit definition |
The amount of enzyme required to dephosphorylate 1 μg of pUC19 DNA digested with HindIII (which produces 5' overhangs) within 30 min at 25℃ is defined as one unit of activity. |
Glycerol Content |
Contains Glycerol |
Storage
The products should be stored at -25~-15℃ for 3 years.
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.
Molecular c loning DNA sequencing SNP analysis. Always verify compatibility with your specific template, buffer, and downstream workflow.
One unit (U) is defined as the amount of enzyme that catalyzes the phosphorylation or dephosphorylation of 1 nmol of substrate per minute at 37°C under standard buffer conditions.
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.
T4 Polynucleotide Kinase (T4 PNK) catalyzes the transfer of a γ-phosphate from ATP to the 5′-hydroxyl terminus of DNA or RNA, enabling ligation of blunt-end or cohesive-end fragments lacking 5′ phosphates. Shrimp Alkaline Phosphatase (SAP) removes 5′ and 3′ phosphate groups from DNA, RNA, and dNTPs, and is heat-inactivated at 65°C for 15 min—eliminating the need for column cleanup prior to downstream reactions.
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.
