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D5025

Sigma-Aldrich

脱氧核糖核酸酶 I 来源于牛胰腺

Type IV, lyophilized powder, ≥2,000 Kunitz units/mg protein

别名:
脱氧核糖核酸 5′-寡核苷酸-水解酶, DNase I
CAS号:
EC 号:
MDL编号:
NACRES:
NA.54

质量水平

300

类型

Type IV

形式

lyophilized powder

specific activity

≥2,000 Kunitz units/mg protein

分子量

~31 kDa

纯化方式

chromatography

组成

Protein, ≥80%

溶解性

0.15 M NaCl: soluble 5.0 mg/mL, clear, colorless

特色行业

Diagnostic Assay Manufacturing
Diagnostic Assay Manufacturing

异质活性

Chymotrypsin ≤0.5%
Protease ≤0.05%
RNase ≤0.02%

运输

wet ice

储存温度

−20°C

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应用

DNAse I可用于切开DNA,作为将标记碱基插入DNA的第一步。来自sigma的该酶已用于处理大鼠的脑组织。该研究表明星形胶质细胞的轴突生长不受少突胶质细胞抑制。在另一项研究中,大肠杆菌的解冻固定样本被来自Sigma的DNAseI以及其他酶进行消化。消化是在进行通透化处理和核酸染色前完成的。
来自牛胰腺的脱氧核糖核酸酶I已用于研究枯草芽孢杆菌中核酸酶的二维酶谱分析。来自牛胰腺的脱氧核糖核酸酶I还用于研究小沟和大沟结合药物和嵌入剂对小沟结合蛋白RecA和脱氧核糖核酸酶I的DNA结合的影响。
用于从蛋白质样品中除去 DNA。

包装

15000 units in glass bottle
150000, 375000, 750000 units in poly bottle

生化/生理作用

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。最适pH在7-8之间。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。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,它们的摩尔比为4:1:1。仅有少量的D被发现。2-巯基乙醇、螯合剂、十二烷基硫酸钠(SDS)和肌动蛋白都已知可抑制酶的活性。

单位定义

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

外形

含氯化钙的冻干粉

制备说明

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。

分析说明

蛋白由双缩脲法测定。

Storage Class Code

13 - Non Combustible Solids

WGK Germany

WGK 3

闪点(F)

Not applicable

闪点(C)

Not applicable

分析证书

原产地证书 (CofO)

  1. How should I prepare a solution of Product D5025, DNAse I?

    This enzyme can be reconstituted in 0.15 M NaCl at a concentration of 10 mg/ml.

  2. Are stock solutions of Product D5025, Deoxyribonuclease I (DNase I), stable?

    Solutions of DNAse I (10 mg/ml) 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. lose approximately 20% activity.

  3. How do Deoxyribonuclease I (DNase I), products D5025 and D4527  differ from each other?

    Both of these products have the same specific activity (>2000 units/mg protein) for Dnase I. The enzyme impurity levels for each, however, are different. D5025 has the following impurity specification: Chymotrypsin (<0.5%), Protease (<0.05%), and RNase (<0.02%). The impurity specifications for D4527 are: Chymotrypsin (<0.01%), Protease (<0.005%), and RNase (<0.01%).

  4. How can I convert units of Product D5025, Deoxyribonuclease I (DNase I), to mass?

    The mg of solid for each unit package size will be indicated on the label of the package. For a particular lot, you can calculate the specific activity in units per mg solid by multiplying the specific activity in units per mg protein by the percent protein divided by 100.

  5. What concentration of Product D5025, Deoxyribonuclease I (DNase I),  is required to remove DNA from solutions?

    DNA can be removed from preparations by incubation with 20 - 50 ug DNAse I in the presence of 50 mM Tris-HCl buffer, pH 7.5, 10 mM magnesium chloride, at 37°C. for 60 min.

  6. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  7. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  8. How do I find price and availability?

    There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers.

  9. What is the Department of Transportation shipping information for this product?

    Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product. 

  10. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

T Guindulain et al.
Applied and environmental microbiology, 63(11), 4608-4611 (1997-11-15)
Three nucleic acid dyes (SYTO-13, TOTO-1, and YOYO-1) were tested on cultures of Escherichia coli and marine prokaryote populations. These dyes stain the RNA and DNA in E. coli but only respond to DNA in marine populations, according to the...
J W Fawcett et al.
Journal of cell science, 103 ( Pt 2), 571-579 (1992-10-01)
Axon growth in vitro may be inhibited by contact with oligodendrocytes, but most axons grow readily on the surface of astrocyte monolayers. Since both cell types are in close contact with one another in the damaged nervous system, we have...
Enzymes of Molecular Biology
Weir, A. F.
Methods in Molecular Biology, 16 (1993)
Chii J Chan et al.
Biophysical journal, 112(6), 1063-1076 (2017-03-30)
Understanding the physical mechanisms governing nuclear mechanics is important as it can impact gene expression and development. However, how cell nuclei respond to external cues such as heat is not well understood. Here, we studied the material properties of isolated...
Sambrook, J., and Russell, D.W.
Molecular Cloning: A Laboratory Manual, 2(2), 5-5 (2001)

实验方案

Enzymatic Assay of Deoxyribonuclease I (EC 3.1.21.1)

To standardize a procedure for the enzymatic assay of Deoxyribonuclease I.

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