Shrimp Alkaline Phosphatase

The main advantages of Shrimp Alkaline Phosphatase (SAP)

  • 100% heat-inactivated at 65°C
  • Removes 5’-phosphates from DNA, RNA, dNTPs and proteins
  • Works in restriction enzyme buffers
  • Used in PCR clean-up

Shrimp Alkaline Phosphatase (SAP) – still the golden standard and the first heat-labile alkaline phosphatase on the market. It is a multipurpose alkaline phosphatase that can be fully inactivated by a short heat treatment.

SAP has been on the market since 1993 and is now replaced by the new and better recombinant form. The Recombinant enzyme is produced in Pichia Pastoris, and has all the properties of the well-proven native version, but with additional benefits. The recombinant version is far more stable at ambient temperature, is also of high, consistent purity, and available at larger batches at high concentration.

 


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Properties

Specifications

Unit definition: One Unit will convert 1 µmol of p-nitrophenyl phosphate per minute to nitrophenol and phosphate at 37ºC and pH 10.4 in 0.1 M glycine buffer, 1 mM each of ZnCl2 and MgCland 6 mM 4-nitrophenyl phosphate.

This Unit definition makes 1 Unit of SAP equivalent to 5 to 40 Units of Antarctic Phosphatase (New England Biolabs), 1.3 Units APex phosphatase or 35 Units NTPhos phosphatase (Epicentre Biotechnologies). SAP is therefore the most processive alternative.

Specific activity: >2 000 Units/mg.
Purity: Purified to apparent homogeneity by SDS-PAGE. DNase, RNase or protease activities not detected.
Concentration: Minimum 10 000 Units/ml, available up to 30 000 Units/ml.
Batch sizes: 25 – 35 million Units / batch.

Stability

Stable at -20ºC in storage buffer (25 mM Tris-HCl pH 7.6, 5 mM MgCl2, glycerol 50%). At room temperature, >95% activity remains after 90 days.

Still, the enzyme is completely inactivated after 5 minutes at 65°C. At 75°C, SAP is completely inactivated after only 1 minute. In a standard thermocycler, the process of heating from 37 to 95 and back to 37°C is sufficient to completely inactivate SAP.

Intellectual property

Recombinant SAP is covered by the following patents: US 7,323,325, EP 1,326,890, JP 4,191,479, AU 9,398,701 and related applications in other countries either granted or pending.

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