NTPhos™ Thermolabile Phosphatase

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Last modified: 2008-07-15 14:57:15

NTPhos™ Thermolabile Phosphatase is a novel, new recombinant enzyme that is highly active in removing phosphate groups from the 5'-position of ribonucleoside- or 2'-deoxyribonucleoside-5'-triphosphates, diphosphates or monophosphates. It can be used to remove 5'-phosphate groups from nucleotide substrates following reactions using enzymes such as RNA-dependent or DNA-dependent RNA or DNA polymerases, terminal transferases or poly(A) polymerases in order to prevent the nucleotides from inhibiting, interfering with, or causing background problems in subsequent downstream reactions.

In other reactions using dye-labeled nucleoside-triphosphates, unincorporated labeled nucleotides can be dephosphorylated following the reaction to prevent background labeling in subsequent reactions. Further, NTPhos™ can be used to remove unused ATP, which is required as an energy source by many enzymes, in order to stop the reaction or assure that the ATP will not intefere in subsequent reactions.

Following removal of 5'-phosphates from the nucleotides, the NTPhos™ enzyme can be completely and irreversibly heat-inactivated by incubation at 65°C for 15 minutes. Since NTPhos™ Phosphatase is manufactured using a recombinant strain, the high quality of NTPhosNTPhos™ Phosphatase is assured and consistent for every lot.

Unit Definition:

One unit results in the release of one nanomole of inorganic phosphate from ATP or dATP in 1 minute at 37°C under standard assay conditions. NTPhos Phosphatase is supplied at a concentration of 20units per microlitre.

Storage Buffer:

50% glycerol containing 50 mM Tris-HCl (pH 7.5), 0.1 mM zinc acetate, 10 mM MgCl2, 0.1 M NaCl, and 0.1% Triton X-100.

Quality Control:

NTPhos Phosphatase is free of detectable DNA exo- and endonuclease and RNase activities.

Guidelines for Nucleotide Hydrolysis
The amount of NTPhos Phosphatase and reaction time depends on the amount of NTP or dNTP substrate to hydrolyze. Use the table below as a guideline to choose the treatment time and amount of enzyme for a given amount of substrate. The entire nucleotide-containing reaction or just an aliquot can be treated directly in most enzyme reaction buffers. Please note that the data in the table was generated using dATP as the substrate in 1X TA buffer*.

Last modified: 2008-05-14 16:46:46

Applications

• Removing 5'-phosphates from modified or unmodified NTPs, NDPs, NMPs, dNTPs, dNDPs, dNMPs, and inorganic pyrophosphate following in vitro reactions using replicases, DNA polymerases, RNA polymerases, terminal transferases, poly(A) polymerases or enzymes that use nucleotide substrates for synthesis, tailing, modification, or labeling of nucleic acids.
• Removing ATP from enzymatic reactions that use it as an energy source.

Benefits

• High Activity - One unit releases 1 nanomole of inorganic phosphate from ATP or dATP in 1 minute at 37°C under standard assay conditions.
• Purified from a recombinant source for consistent high quality and performance.
• Quick & Easy Inactivation - NTPhos™ Phosphatase is completely and irreversibly heat-inactivated in 15 minutes at 65°C.
• Active in RNA and DNA Polymerase & Replicase Buffers - NTPhos™ Phosphatase is active in a wide range of buffers at pH 7-10, and in the presence of 1 M Na+, NH4+, K+, Cl- or acetate- or 10% Triton X-100.

Last modified: 2008-05-14 16:46:46

Protocols for: NTPhos™ Thermolabile Phosphatase

(catalogue number NT4905H / NT4910K / NT4920K)

Please note: all protocols off site are the responsibility of the products supplier

Last modified: 2008-05-14 16:46:46