Antarctic Phosphatase

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
M0289S     -20       Antarctic Phosphatase M0289SVIAL -20 1 x 0.2 ml 5,000 units/ml   Antarctic Phosphatase Reaction Buffer B0289SVIAL -20 1 x 1.5 ml 10 X
M0289L     -20       Antarctic Phosphatase M0289LVIAL -20 1 x 1 ml 5,000 units/ml   Antarctic Phosphatase Reaction Buffer B0289SVIAL -20 1 x 1.5 ml 10 X

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.
• Treatment of dNTPs in PCR reactions prior to sequencing or SNP analysis.

Companion Products

• T4 DNA Ligase
• Quick Ligation™ Kit
• Instant Sticky-end Ligase Master Mix
• Blunt/TA Ligase Master Mix
• Monarch® Plasmid Miniprep Kit 
• Monarch® DNA Gel Extraction Kit
• Monarch® PCR & DNA Cleanup Kit (5 μg)
• Antarctic Phosphatase Reaction Buffer

1. Adding 1/10 volume of 10X Antarctic Phosphatase Reaction Buffer will provide the amount of Zn²⁺ that the enzyme requires for activity.
2. Antarctic Phosphatase is also active in all restriction enzyme NEBuffers 1.1, 2.1, 3.1 and CutSmart^® Buffer only when supplemented with 1/10 volume of the 10X Antarctic Phosphatase Reaction Buffer.
3. Antarctic Phosphatase activity is enhanced in the presence of monovalent salts.
4. Antarctic Phosphatase is inhibited by metal chelators (e.g. EDTA), inorganic phosphate and phosphate analogs. Antarctic Phosphatase activity is decreased in the presence of reducing agents (DTT, β-ME).
5. Molecular Weight: Antarctic Phosphatase is a homodimer. The molecular weight of the monomer is 35 kDa.
6. Antarctic Phosphatase, as are most alkaline phosphatases, is a Zn²⁺ and Mg²⁺ –dependent enzyme and does require supplemental zinc.

1. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual,
   . (2nd ed.), 5.72.
2. Rina, M. et al. (2000). Eur. J. Biochem.. 267, 1230-1238.
3. Guthrie, E., unpublished results. Unpublished observation

1. Protocol for Dephosphorylation of 5´-ends of DNA using Antarctic Phosphatase (NEB #M0289)

• Activity of DNA Modifying Enzymes in rCutSmart™ Buffer

• NEB Diluent and Buffer Table
• Phosphatase Selection Chart

• NEBcloner^®

1. Which alkaline phosphatase, Quick CIP, rSAP or Antarctic Phosphatase works best?
2. The number of colonies that don't contain an insert seems high, how can I tell if the Antarctic Phosphatase worked?
3. Will Quick CIP, rSAP or Antarctic Phosphatase (AnP) dephosphorylate proteins?
4. Does the DNA need to be purified after the restriction digest, prior to Antarctic Phosphatase treatment?
5. Does the DNA need to be purified after Antarctic Phosphatase treatment?
6. What is the molecular weight?
7. Can Antarctic Phosphatase be heat inactivated?
8. What phosphate groups are removed by rSAP, Quick CIP, or Antarctic Phosphatase (AnP)?
9. Does the DNA need to be purified after a restriction digest and prior to the dephosphorylation step?
10. Does the DNA need to be purified after the dephosphorylation step and prior to the ligation step?
11. Are the alkaline phosphatases active in NEBuffers?
12. Cloning Problem: Too much background in the transformation step.

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.

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]

• M0289S_L_v2

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]

• M0289S_L_v1_0221603
• M0289S_L_v1_0351611
• M0289S_L_v1_0221504
• M0289S_L_v1_0371702
• M0289S_L_v1_0371710
• M0289L_v1_10014915
• M0289S_v1_10014916
• M0289S_v1_10027177
• M0289L_v1_10034941
• M0289S_v1_10034943
• M0289L_v1_10040032
• M0289S_v1_10044638
• M0289S_v1_10052532
• M0289L_v1_10052530
• M0289S_v2_10056361
• M0289L_v2_10068198
• M0289S_v2_10067113
• M0289L_v2_10070698
• M0289S_v2_10068197
• M0289L_v2_10077395
• M0289S_v2_10075877
• M0289S_v2_10090704
• M0289L_v2_10087266
• M0289S_v2_10095245
• M0289S_v2_10100262
• M0289L_v2_10124289
• M0289S_v2_10132856
• M0289S_v2_10116043
• M0289S_v2_10140301
• M0289L_v2_10144345
• M0289S_v2_10144347
• M0289S_v2_10157348
• M0289L_v2_10162297
• M0289S_v2_10162300
• M0289S_v2_10185554
• M0289L_v2_10208059
• M0289L_v2_10215907
• M0289S_v2_10225766
• M0289S_v2_10239503
• M0289L_v2_10236898
• M0289S_v2_10251935

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• Antarctic Phosphatase
• Antarctic Phosphatase Reaction Buffer

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.