New England Biolabs Canada
Expand menu
Sign In or Sign UpMy NEB

Antarctic Phosphatase

Catalog # Concentration Size List Price Quantity Your Price
M0289L 5000 units/ml 5000 units $477.00
$429.30
M0289S 5000 units/ml 1000 units $117.00
$105.30
Catalog # Size List Price Your Price
M0289L 5000 units $477.00
$429.30
M0289S 1000 units $117.00
$105.30
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

Antarctic Phosphatase (AnP) is a heat labile alkaline phosphatase purified from a recombinant source.

  • Rapid and irreversible heat inactivation eliminates unwanted activity
  • Flexible reaction conditions (active in any restriction enzyme buffer, no clean-up required)
  • No need for multiple phosphatases (AnP removes 5´- and 3´- phosphates from DNA, RNA and dNTPs)
  • Active on unincorporated dNTPs in PCR products - improves DNA sequencing and SNP analysis

Featured Videos
View Video Library
  • Monarch_MiniprepProtocol_VideoThumb
    The Mechanism of DNA Phosphorylation

Antarctic Phosphatase (AnP) is a heat labile alkaline phosphatase purified from a recombinant source. AnP nonspecifically catalyzes the dephosphorylation of 5´ and 3´ ends of DNA and RNA phosphomonoesters. Also, AnP Hydrolyses ribo-, as well as deoxyribonucleoside triphosphates (NTPs and dNTPs). AnP 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. AnP may also be used to degrade unincorporated dNTPs in PCR reactions to prepare templates for DNA sequencing or SNP analysis. AnP is completely and irreversibly inactivated by heating at 80°C for 2 minutes, thereby making removal of AnP prior to ligation or end-labeling unnecessary. 

Product Source
An E. coli strain that carries the TAB5 AP gene, originally cloned in plasmid pNI (2), recloned in plasmid pEGTAB7-4.1(3).
Reagents Supplied

The following reagents are supplied with this product:

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

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme that will dephosphorylate 1 µg of pUC19 vector DNA cut with a restriction enzyme generating 5´ recessed ends in 30 minutes at 37°C. Dephosphorylation is defined as > 95% inhibition of recircularization in a self-ligation reaction and is measured by transformation into E. coli.

Reaction Conditions

1X Antarctic Phosphatase Reaction Buffer
Incubate at 37°C

1X Antarctic Phosphatase Reaction Buffer
50 mM Bis-Tris-Propane-HCl
1 mM MgCl2
0.1 mM ZnCl2
(pH 6 @ 25°C)

Storage Buffer

10 mM Tris-HCl
1 mM MgCl2
50% Glycerol
0.01 mM ZnCl2
pH 7.4 @ 25°C

Heat Inactivation

80°C for 2 minutes

Molecular Weights

Apparent: 35 kDa
Theoretical: 69 kDa

Unit Assay Conditions

Vector DNA is dephosphorylated in restriction endonuclease buffer supplemented with Antarctic Phosphatase Reaction Buffer. Ligation is performed with 50 ng of vector using the NEB Quick Ligation Kit (NEB #M2200).

Companion Products
Notes
  • Adding 1/10 volume of 10X Antarctic Phosphatase Reaction Buffer will provide the amount of Zn2+ that the enzyme requires for activity.
  • 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.
  • Antarctic Phosphatase activity is enhanced in the presence of monovalent salts.
  • 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).
  • Molecular Weight: Antarctic Phosphatase is a homodimer. The molecular weight of the monomer is 35 kDa.
  • Antarctic Phosphatase, as are most alkaline phosphatases, is a Zn2+ and Mg2+ –dependent enzyme and does require supplemental zinc.
References
  • Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual,
    . (2nd ed.), 5.72.
  • Rina, M. et al. (2000). Eur. J. Biochem.. 267, 1230-1238.
  • Guthrie, E., unpublished results. Unpublished observation
Quality Control Assay
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
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]
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]
Safety Data Sheets
Legal And Disclaimer

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.

Other Products You May Be Interested In
 Top