General description
DNase I, or Deoxyribonuclease I, is an endonuclease isolated from bovine pancreas.
• Our DNase I Digests double str and single stranded DNA into oligo and mononucleotides.
• Bovine pancreatic DNase exists as four isozymes, having isoelectric points for A, B, C and D: 5.22, 4.96, 5.06 and 4.78.3. The predominant form is A, with smaller amounts of B and C, and only minor amount of D.
• DNase I structure resembles the structure of to exonuclease III. It includes two central ß sheets. Each β sheet is composed of six β-strands. This complex of β sheets is surrounded by extensive loop and α-helical regions. This enzyme shares structural similarity to exonuclease III.[1]
Application
• Decreases viscosity providing better yields by removing DNA in primary cell isolation:
• Incorporating labelled bases into DNA: DNA nick
• Radioactive labelling
• Bioprocessing applications: DNA removal
• Eliminating genomic DNA from RNA preparations before RT-PCR
• In vitro transcription
• Nick translation
• DNase footprinting
• Actin regulation of actin polymerization in cells, and cell apoptosis
• UV crosslinking of proteins to nucleic acids
• DNase play a role in the regulation of actin polymerization in cells and is involved in apoptosis process [1]
用于从蛋白质样品中除去 DNA。
包装
1, 5 g in poly bottle
10, 100 mg in poly bottle
Biochem/physiol Actions
DNase I是一种内切核酸酶,可作用于与嘧啶相邻的磷酸二酯键以产生具有末端5′-磷酸的聚核苷酸。当Mg2+存在时,DNAse I可独立切割DNA的每条链且切割位点是随机的。当Mn2+存在时,两条DNA链都几乎是在同样的位置被切割。[6]二价阳离子如Mn2+, Ca2+, Co2+以及Zn2+都是酶的激活剂。5 mM浓度的Ca2+可稳定酶免收蛋白水解酶的消化。最适pH在7-8之间。[7]来自牛胰腺的DNAse I由四种色谱可区分的组分A,B,C和D组成,它们的摩尔比分别为4:1:1。仅发现少量D。[5]2-巯基乙醇、螯合剂、十二烷基硫酸钠(SDS)[4]和肌动蛋白[1]都已知可抑制酶的活性。Features and Benefits
Our Deoxyribonuclease DNAse I,is essentially RNAse free product, which support the product application.
Unit Definition
一Kunitz单位的酶在以I型或III型DNA作为底物时,在 pH 5.0,37°C的条件下在每mL中每分钟可引起0.001的A260改变。Physical form
粗制品,含有氯化钙
Preparation Note
溶液0.15 M NaCl的10 mg/mL DNAse I溶液分装保存在−20 °C一周后可能会丢失<10%的活性。同样的溶液保存在2-8 °C则可能丢失约20%的活性。当在pH 5和7之间时,它可在60 °C维持活性最长达五小时,并在68 °C <10分钟内丢失活性。它在乙酸缓冲液(pH 5.0)和tris缓冲液((pH 7.2),1 mg/mL浓度)中以6%/小时的速率丢失活性。Analysis Note
Protein determined by biuret.