产品名称

Deoxyribonuclease I from bovine pancreas

别 称

DNase I, Deoxyribonucleate 5′-oligonucleotido-hydrolase

含 量

Standardized vial containing 2,000 Kunitz  units of DNase I (D4527), vial of ≥0.25 mg total protein

CAS  NO.

9003-98-9

分子量

mol wt ~31 kDa

MDL  number

MFCD00130918

适用范围

生物试剂

l  理化信息

外 观

白色至浅黄色粉末

溶解性(25℃)

0.15 M NaCl: soluble 5.0 mg/mL, clear

l  生物学信息

生物活性/药理作用

DNase I is an endonuclease that acts on phosphodiester bonds adjacent to pyrimidines to produce polynucleotides with terminal 5′-phosphates. In the presence of Mg₂₊, DNAse I cleaves  each strand of DNA independently and the cleavage sites are random. Both DNA strands are cleaved at approximately the same site in the presence of Mn₂₊. The pH optimum is found to be between 7 and 8. Divalent cations such as Mn₂₊, Ca₂₊, Co₂₊, and Zn₂₊ are activators of the enzyme. A concentration of 5 mM Ca₂₊ stabilizes the enzyme against proteolytic digestion. DNAse I from bovine pancreas consists of four chromatographically distinguishable components, A, B, C, and D, with molar ratios being 4:1:1 respectively. Only minor amounts of D are found. 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS) and actin are known to inhibit the enzyme activity.

应 用

Used for the removal of DNA from protein samples.

DNAse I is used to nick DNA as a first step to incorporate labeled s into DNA. DNAse I from Sigma has been used along with other enzymes for tumor harvest and dissociation, during the isolation and molecular characterization of cancer stem cells in MMTV-Wnt-1 murine breast tumors.

Deoxyribonuclease I from bovine pancreas has been used in a study to determine that mammalian deoxyribonucleases I are classified into three types d on differences in their tissue concentrations.Deoxyribonuclease I from bovine pancreas has also been used in a study to compare the three primary structures of deoxyribonuclease isolated from bovine,ovine, and porcine pancreas.

样品准备说明

The enzyme powder may be reconstituted in water or any buffer at pH 4.0-9.0, except phosphate buffer. Calcium chelators  should be avoided. 10mg/mL solution of DNAse I in 0.15M NaCl may lose <10% of its activity when stored for a week in aliquots at −20 °C. The same solutions stored in aliquots at 2-8 °C can lose approximately 20% activity. It remains active for up to five hours at 60 °C between pH 5 and 7, and loses activity in <10 minutes at 68°C. It loses activity at the rate of 6%/hour in acetate buffer (pH5.0) and tris buffer ((pH7.2) at 1 mg/mL concentration.

有效单位定义

One Kunitz unit will produce a ΔA₂₆₀ of 0.001 per min per mL at pH5.0 at 25°C, using DNA, Type I or III as substrate.

l  存储

存储温度

-20°C

l  注意事项及免责声明

本产品仅用于实验研究，不得作为药物使用，不得用于家用或其它用途。

l  参考文献

1.    http://www.drugbank.ca

2.    https://ncit.nci.nih.gov

3.    https://www.ncbi.nlm.nih.gov