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Home DNA Modifying Enzymes and Cloning Technologies Products Shrimp Alkaline Phosphatase (rSAP)

Shrimp Alkaline Phosphatase (rSAP)

We are excited to announce that all reaction buffers are now BSA-free. NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Find more details at www.neb.com/BSA-free.

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

Note: See also Exo-CIP™ Rapid PCR Cleanup Kit (NEB #E1050).

Catalog #	Concentration	Size	List Price	Your Price
M0371S	1,000 units/ml	500 units	$64.00	Sign In Sign In
M0371L	1,000 units/ml	2,500 units	$258.00	Sign In Sign In

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Product Information

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 Source

A Pichia pastoris clone that carries the shrimp alkaline phosphatase gene from Northern shrimp Pandalus borealis (1,2).

This product is related to the following categories:: Phosphatases & Sulfurylases Products,; RNA Modification Products

This product can be used in the following applications:: Dephosphorylation,; RNA Modification,; In vitro Transcription for RNA Synthesis

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

M0371S -20

Shrimp Alkaline Phosphatase (rSAP) M0371SVIAL -20 1 x 0.5 ml 1,000 units/ml

rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X

M0371L -20

Shrimp Alkaline Phosphatase (rSAP) M0371LVIAL -20 2 x 1.25 ml 1,000 units/ml

rCutSmart™ Buffer B6004SVIAL -20 2 x 1.25 ml 10 X

Properties & Usage

Unit Definition
One unit is defined as the amount of enzyme that hydrolyzes 1 μmol of p-Nitrophenyl Phosphate, PNPP (NEB #P0757) in a total reaction volume of 1 ml in 1 minute at 37°C.
Reaction Conditions

1X rCutSmart™ Buffer
Incubate at 37°C

1X rCutSmart™ Buffer
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Storage Buffer

25 mM Tris-HCl
1 mM MgCl₂
50% Glycerol
pH 7.5 @ 25°C

Heat Inactivation
65°C for 5 minutes
Molecular Weight
Theoretical: 54 kDa

Unit Assay Conditions
1 M diethanolamine-HCl (pH 9.8), 0.5 mM MgCl₂, 50 mM p-Nitrophenyl Phosphate. These conditions are only used for quantitating enzyme activity.
Advantages and Features
Application Features
- Dephosphorylation 5´ and 3´ ends of DNA and RNA.
- Dephosphorylation of cloning vector DNA to prevent recircularization during ligation.
- Dephosphorylation of DNA prior to end-labeling using T4 Polynucleotide Kinase.
- Removal of dNTPs in PCR reactions prior to sequencing or SNP analysis.
Related Products
Companion Products
Product Notes
1. rSAP, as are most alkaline phosphatases, is a Zn²⁺ and Mg²⁺-dependent enzyme. Our formulation has tightly bound zinc atoms in the active center and does not require supplemental zinc or other additives.
2. rSAP is also active in 1X NEBuffers 1.1, 2.1, 3.1 as well as NEBuffers 1, 2, 3, 4 and NEBuffer for EcoRI.
3. rSAP activity is enhanced in the presence of monovalent salts.
4. rSAP is inhibited by metal chelators (e.g. EDTA), inorganic phosphate and phosphate analogs.
5. The rSAP activity is decreased in the presence of reducing agents (DTT, β-ME).
6. Molecular Weight: rSAP is a homodimer. Themolecular weight of the monomer is 54 kDa.
References
1. Olsen, R. L. et al. (1991). Comp. Biochem. Physiol. 99B(4):, 755-761.
2. Nilsen, I.W. et al. (2001). Comp. Biochem. Physiol. 129B(4):, 853-861.

Protocols, Manuals & Usage
Tools & Resources
- Selection Charts
  NEB Diluent and Buffer Table
  
  Phosphatase Selection Chart
- Web Tools
  NEBcloner®
FAQs & Troubleshooting
- FAQs
  Which alkaline phosphatase, Quick CIP, rSAP or Antarctic Phosphatase works best?
  
  The number of colonies that do not contain an insert seems high. How can I tell if rSAP worked?
  
  What phosphate groups are removed by rSAP, Quick CIP, or Antarctic Phosphatase (AnP)?
  
  Does the DNA need to be purified after rSAP treatment?
  
  How stable is rSAP in its storage buffer at various temperatures?
  
  What is the effect of metal chelators, inorganic phosphate and phosphate analogs on rSAP activity?
  
  What is the effect of reducing agents on rSAP activity?
  
  Will Quick CIP, rSAP or Antarctic Phosphatase (AnP) dephosphorylate proteins?
  
  Can rSAP be heat inactivated?
  
  Does the DNA need to be purified after a restriction digest and prior to the dephosphorylation step?
  
  Does the DNA need to be purified after the dephosphorylation step and prior to the ligation step?
  
  Are the alkaline phosphatases active in NEBuffers?
  
  Cloning Problem: Too much background in the transformation step.
  
  Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
Citations & Technical Literature
- Citations
  
  Product Citation Tool
  
  Powered by Bioz See more details on Bioz
Quality, Safety & Legal
- Quality Control Assays
  
  Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
- Specifications & Change Notifications
  Specifications & Change Notifications
  The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
  
  M0371S_L_v1
- Certificate of Analysis
  The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
  
  M0371S_L_v1_0031607
  
  M0371S_L_v1_0031608
  
  M0371S_L_v1_0031612
  
  M0371S_L_v1_0041701
  
  M0371S_L_v1_0041709
  
  M0371S_L_v1_0051711
  
  M0371S_v1_10008070
  
  M0371S_v1_10018074
  
  M0371L_v1_10008068
  
  M0371S_v1_10030296
  
  M0371L_v1_10030298
  
  M0371S_v1_10041601
  
  M0371L_v1_10041602
  
  M0371L_v1_10048761
  
  M0371S_v1_10048764
  
  M0371L_v1_10056272
  
  M0371L_v1_10057228
  
  M0371S_v1_10057229
  
  M0371S_v1_10064355
  
  M0371L_v1_10064354
  
  M0371S_v1_10069213
  
  M0371S_v1_10081111
  
  M0371S_v1_10085490
  
  M0371L_v1_10091812
  
  M0371S_v1_10091811
  
  M0371S_v1_10098689
  
  M0371L_v1_10106617
  
  M0371S_v1_10111319
  
  M0371S_v1_10121444
  
  M0371L_v1_10127370
  
  M0371L_v1_10134722
  
  M0371S_v1_10134720
  
  M0371S_v1_10143863
  
  M0371S_v1_10153699
  
  M0371L_v1_10153911
  
  M0371S_v1_10153913
  
  M0371S_v1_10168761
  
  M0371S_v1_10185192
- Safety Data Sheets
  The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
  
  Shrimp Alkaline Phosphatase (rSAP)
  
  rCutSmart™ Buffer
- Legal and Disclaimers
  
  Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.
  
  This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
  
  New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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Shrimp Alkaline Phosphatase (rSAP)

Figure 1: rSAP heat inactivation at 65°C

Product Source

Reagents Supplied

Unit Definition

Reaction Conditions

Storage Buffer

Heat Inactivation

Molecular Weight

Unit Assay Conditions

Application Features

Companion Products

Product Citation Tool

Specifications & Change Notifications

Other Products You May Be Interested In

Monarch^® PCR & DNA Cleanup Kit (5 μg)

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Shrimp Alkaline Phosphatase (rSAP)

Featured Video

The Mechanism of Dephosphorylation

Figure 1: rSAP heat inactivation at 65°C

Product Source

Reagents Supplied

Unit Definition

Reaction Conditions

Storage Buffer

Heat Inactivation

Molecular Weight

Unit Assay Conditions

Application Features

Companion Products

Product Citation Tool

Specifications & Change Notifications

Featured Videos

The Mechanism of Dephosphorylation

Other Products You May Be Interested In

Monarch® PCR & DNA Cleanup Kit (5 μg)

Blunt/TA Ligase Master Mix

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