Type II, lyophilized powder, Protein ≥80 %, ≥2,000 units/mg protein

别名: DNase I, 脱氧核糖核酸 5′-寡核苷酸-水解酶

• CAS号 9003-98-9

•  Enzyme Commission (EC) Number 3.1.21.1  ( BRENDA | IUBMB )

•  EINECS 编号 232-667-0

•  MDL number MFCD00130918
  
  

  性质

  Related Categories	3.1.x.x Acting on esters, 3.x.x.x Hydrolases, Application Index, Enzyme Class Index, Enzymes for Diagnostic Kit Manufacturing, General Metabolic Enzymes, Molecular Diagnostics, Nucleotide Metabolism, Protein & Nucleic Acid Isolation, 代谢组学, 分子生物学, 天然蛋白质样品制备, 生化试剂, 细胞生物学, 细胞裂解和蛋白质提取试剂, 蛋白样本净化, 蛋白质组学, 酶、抑制剂和底物 更多...
  Quality Level	PREMIUM
  type	Type II
  form	lyophilized powder
  mol wt	mol wt ~31 kDa
  purified by	chromatography
  composition	Protein, ≥80%
  solubility	0.15 M NaCl: soluble 5.0 mg/mL, clear
  Featured Industry	Diagnostic Assay Manufacturing Diagnostic Assay Manufacturing
  foreign activity	Chymotrypsin ≤0.01%
  	Protease ≤0.005%
  	RNase ≤0.01%
  shipped in	wet ice
  storage temp.	−20°C

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  产品描述

  Analysis Note

  Protein determined by biuret.

  Application

  用于从蛋白质样品中除去 DNA。

  DNAse I is used to nick DNA, as a first step to incorporate labeled bases into DNA. The enzyme from Sigma has been used in the nuclease stock along with RNAse during cell lysate preparation of Madin-Darby canine kidney (MDCK) II cell lines.^[2] It has also been used during RNA extraction from Sindbis virus.^[3]

  Deoxyribonuclease I from bovine pancreas has been used for the identification, localization ａnd expression of two novel human genes similar to deoxyribonuclease I. Deoxyribonuclease I from bovine pancreas has also been used in a study to investigate a new approach to obtaining high-activity RNase, DNase, cholesterolesterase, ａnd trypsin from cattle pancreas.

  包装

  10000 units in glass bottle

  20000, 40000, 200000, 500000 units in poly bottle

  Preparation Note

  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. 10 mg/mL solution of DNAse I in 0.15 M 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 (pH 5.0) ａnd tris buffer ((pH 7.2) at 1 mg/mL concentration.

  Unit Definition

  One Kunitz unit will produce a change in A₂₆₀ of 0.001 per minute per ml at pH 5.0 at 25 °C using DNA, Type I or III, as the substrate.

  Physical form

  Contains calcium chloride

  Biochem/physiol Actions

  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⁺.^[6] 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.^[7] 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 their molar ratios being 4:1:1 respectively. Only minor amounts of D are found.^[5] 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS)^[4] ａnd actin^[1] are known to inhibit the enzyme activity.