应用领域
NTPhos™ Thermolabile Phosphatase is a novel 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.
Following removal of 5´ phosphates from nucleotides, the NTPhos enzyme can be completely and irreversibly heat-inactivated by incubation at 65°C for 15 minutes.
NTPhos™不耐热磷酸酶是一种新型的重组酶，对从核糖核苷或2´-脱氧核糖核苷5´-三磷酸，二磷酸或一磷酸的5´位置去除磷酸基团具有高活性。反应后，可以使用它（例如依赖于RNA或依赖于DNA的RNA或DNA聚合酶，末端转移酶或poly（A）聚合酶）从核苷酸底物上除去5´-磷酸基团，以防止核苷酸被抑制，在随后的下游反应中干扰或引起背景问题。
从核苷酸中去除5´磷酸后，可以在65°C孵育15分钟，将NTPhos酶完全不可逆地热灭活。
参数说明
Unit Definition: One unit of NTPhos Thermolabile Phosphatase results in the release of 1 nmol of inorganic phosphate from ATP or dATP in 1 minute at 37°C under standard assay conditions.
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.
单位定义：在标准测定条件下，在37°C下，一分钟NTPhos不耐热磷酸酶可在1分钟内从ATP或dATP中释放出1 nmol的无机磷酸盐。
储存缓冲液：含有50mM的Tris-HCl（pH为7.5），0.1mM的醋酸锌，10毫摩尔MgCl 50％甘油2，0.1M NaCl和0.1％的Triton X-100。
质量控制： NTPhos磷酸酶不含可检测的DNA外切核酸酶和内切核酸酶以及RNase活性。