所有图片(1)

04716728001

Roche

重组DNase I,无RNase

from bovine pancreas, expressed in Pichia pastoris

质量水平

100

生物来源

bovine pancreas

重组

expressed in Pichia pastoris

形式

solution

分子量

~39 kDa

包装

pkg of 10,000 U

optimum pH

7.0-8.0

运输

dry ice

一般描述

重组DNase I是一种DNA特异性内切酶。该酶可通过水解连接到DNA上的磷酸二酯而催化双链及单链DNA的随机降解,形成寡核苷酸和单核苷酸的混合物。重组DNase I的生产过程中所使用的所有材料都无动物来源,因而可形成无动物源产物。

内容物
  • 重组DNasae I,不无RNase,10 U/μl
  • 孵育缓冲液,10x浓度

特异性

热灭活:一单位无RNase重组DNase I可在75 °C孵育10分钟进行热灭活。

重要提示:无RNase重组DNase I也可根据标准实验方案通过酚提取进行灭活并去除,分子生物学中的现有实验方案。

应用

在对RNase较为敏感的应用中,可采用无RNase的重组DNase I降解DNA。例如,DNase I常用于:
  • 在RT-PCR之前去除RNA中的基因组DNA
  • 体外转录反应之后分离不含DNA的RNA
  • 进行缺口翻译
  • 在真核DNA中定位DNase敏感的区域

特点和优势

  • 可从任意RNA样品中去除DNA污染
  • 未检测出RNase或蛋白酶活性
  • 可热灭活,因而免去了有机提取的必要
  • 通过优化的孵育缓冲液进行运输,以最大限度维持DNase活性
  • 完全无动物来源过程生产,去除了动物来源材料相关的任何风险

包装

1个试剂盒包含2种组分

质量

杜绝污染:每批次都按照现有质检工艺检测以确保无RNase及蛋白酶。

产品规格

糖基化形式
重组DNase I异质性N端糖基化,因此会在电泳时呈现两条带。
二价离子要求
DNase I需要二价阳离子以实现最高活性。DNA特异性内切酶通过镁离子等离子激活,并由钙离子刺激。因此,酶会受到EDTA等金属螯合剂抑制。

单位定义

一单位指的在1ml的检测条件下可造成260 nm处吸光值提高0.001/min的酶活性。
反应条件:
根据以下检测混合物测定体积活性。将100 μg小牛胸腺DNA与40至70单位的无RNase重组DNase I在1x孵育缓冲液中+ 25°C下孵育。测定260nm处吸光度增量。

制备说明

激活剂:二价金属离子
工作溶液:储备溶液:20 mM Tris-HCl,50 mM NaCl,2 mM CaCl2,2 mM MgCl2,1mM二硫赤藓糖醇,0.1 mg/ml Pefabloc SC,50% 甘油(v/v),pH 7.6 (4 °C)。
孵育缓冲液(10x):400 mM Tris-HCl,100 mM NaCl,60 mM MgCl2 10 mM CaCl2,pH 7.9。
酶稀释液:25 mM Tris-HCl,50%甘油(v/v),pH 7.6 (4 °C)。

储存及稳定性

将未稀释的酶溶液-15至-25°C储存;缓冲液4°C储存。

其他说明

仅用于生命科学研究。不可用于诊断。

Storage Class Code

12 - Non Combustible Liquids

WGK Germany

WGK 1

闪点(F)

does not flash

闪点(C)

does not flash

分析证书

原产地证书 (CofO)

Kwok-Fong Chan et al.
Molecules (Basel, Switzerland), 25(24) (2020-12-17)
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Kirsten Richter et al.
The Journal of biological chemistry, 295(23), 7849-7864 (2020-04-23)
Activation of the T cell receptor (TCR) results in binding of the adapter protein Nck (noncatalytic region of tyrosine kinase) to the CD3ϵ subunit of the TCR. The interaction was suggested to be important for the amplification of TCR signals...
Christina N Cheng et al.
Journal of visualized experiments : JoVE, (89)(89), doi:10-doi:10 (2014-08-01)
The zebrafish embryo is now commonly used for basic and biomedical research to investigate the genetic control of developmental processes and to model congenital abnormalities. During the first day of life, the zebrafish embryo progresses through many developmental stages including...
Lucas R Smith et al.
Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 54(3), 333-353 (2020-04-11)
Cell migration and extracellular matrix remodeling underlie normal mammalian development and growth as well as pathologic tumor invasion. Skeletal muscle is no exception, where satellite cell migration replenishes nuclear content in damaged tissue and extracellular matrix reforms during regeneration. A...
In Kyoung Mah et al.
Developmental biology, 416(1), 82-97 (2016-06-18)
Polarized epithelia define boundaries, spaces, and cavities within organisms. Cavitation, a process by which multicellular hollow balls or tubes are produced, is typically associated with the formation of organized epithelia. In order for these epithelial layers to form, cells must...

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实验方案

DNase I Recombinant, RNase-free Protocol

DNase I from bovine pancreas is a glycoprotein of Mr 37000. A special procedure is used to remove RNases from the DNase preparation.

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