Shrimp Alkaline Phosphatase (rSAP)
Shrimp Alkaline Phosphatase (rSAP) is a heat labile alkaline phosphatase purified from a recombinant source.
- Rapid and irreversible heat inactivation eliminates unwanted activity
- Improved storage stability versus native enzyme
- Faster reaction setup (no supplemental additives like zinc required)
- Flexible reaction conditions (active in any restriction enzyme buffer, no clean-up required)
- Less enzyme required (high specific activity), resulting in a lower cost per reaction
- No need for multiple phosphatases (rSAP removes 5´- and 3´- phosphates from DNA, RNA and dNTPs )
- Active on unincorporated dNTPs in PCR products - improves DNA sequencing and SNP analysis
- Recombinant for purity, consistency and value
The Mechanism of Dephosphorylation
Shrimp Alkaline Phosphatase (rSAP) is a heat labile alkaline phosphatase purified from a recombinant source. rSAP is identical to the native enzyme and contains no affinity tags or other modifications. rSAP nonspecifically catalyzes the dephosphorylation of 5´ and 3´ ends of DNA and RNA phosphomonoesters. Also, rSAP hydrolyses ribo-, as well as deoxyribonucleoside triphosphates (NTPs and dNTPs). rSAP is useful in many molecular biology applications such as the removal of phosphorylated ends of DNA and RNA for subsequent use in cloning or end-labeling of probes. In cloning, dephosphorylation prevents religation of linearized plasmid DNA. The enzyme acts on 5´ protruding, 5´ recessed and blunt ends. rSAP may also be used to degrade unincorporated dNTPs in PCR reactions to prepare templates for DNA sequencing or SNP analysis. rSAP is completely and irreversibly inactivated by heating at 65°C for 5 minutes, thereby making removal of rSAP prior to ligation or end-labeling unnecessary.
1 unit of rSAP was incubated under recommended reaction conditions, including DNA, for 30 minutes and then heated at 65°C. Remaining phosphatase activity was measured by PNPP assay.
Product SourceA Pichia pastoris clone that carries the shrimp alkaline phosphatase gene from Northern shrimp Pandalus borealis (1,2).
The following reagents are supplied with this product:
Store at (°C) Concentration CutSmart® Buffer -20 10X
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