末端转移酶(小牛胸腺) 制备方法及用途

末端转移酶(小牛胸腺) 物化性质

存储条件/存储方法：

充氩-20°C密闭，避光，通风干燥处

稳定性相关：

Analysis Note Activity assay: 200 mM potassium cacodylate, pH 7.2, 4 mM MgCl2, 1 mM 2-mercaptoethanol, 1 mM 3H-dATP, 70 μM d(pT)6, 37 °C.

Biochem/physiol Actions Bovine terminal transferase (TdT) is a primer-dependent polymerase that catalyzes the addition of deoxynucleotides to the 3′-OH terminus of DNA molecules with the release of inorganic phosphate. TdT reacts preferentially with either single-stranded DNA molecules or double-stranded-DNA with 3′ overhangs, but procedures have been developed to label blunt ends or 3′-recessive ends. In a reaction mixture, the divalent ion (Co2+, Mn2+, Mg2+) will influence purine and pyrimidine polymerization rate. Activities of TdT are also affected by the bases (dATP, dCTP, dGTP and dTTP) present. Each reaction mixture needs to be optimized. The enzyme is commonly used to add complementary homopolymer tails to vector and insert DNA for construction of recombinant DNA in vitro.

Features and Benefits • Add homopolymers to vectors, inserts and cDNA for cloning

• Label the 3′-end of double- and single-stranded DNA with non-radioactive and radioactive labels

• Carry out in vitro mutagenesis by adding single nucleotides to DNA

• Use in TUNEL assays with deoxyuridine triphosphate (dUTP) conjugated with a fluorescent chromophore or biotin to label nicked DNA in apoptotic cells

 

Unit Definition One unit will incorporate 1 nanomole of dATP into acid-precipitable material in one hour at 37 °C using d(pT)6 as primer.

Physical form Solution in 50 mM potassium phosphate, pH 7.4, 1 mM 2-mercaptoethanol and 50% glycerol (v/v)

其它信息：

1.      性状：冷冻干燥含氯化钾的固体。

2.       密度（g/mL,25℃）：未确定

3.       相对蒸汽密度（g/mL,空气=1）：未确定

4.       熔点（ºC）：未确定

5.       沸点（ºC,常压）：未确定

6.       沸点（ºC,5.2kPa）：未确定

7.       折射率：未确定

8.       闪点（ºC）：未确定

9.       比旋光度（º）：未确定

10.    自燃点或引燃温度（ºC）：未确定

11.    蒸气压（kPa,20ºC）：未确定

12.    饱和蒸气压（kPa,60ºC）：未确定

13.    燃烧热（KJ/mol）：未确定

14.    临界温度（ºC）：未确定

15.    临界压力（KPa）：未确定

16.    油水（辛醇/水）分配系数的对数值：未确定

17.    爆炸上限（%,V/V）：未确定

18.    爆炸下限（%,V/V）：未确定

19.    溶解性：未确定

末端转移酶(小牛胸腺) 毒理性