DNase I from bovine pancreas

DNase I

from bovine pancreas

General description

DNase I is a double-strand specific endonuclease that degrades DNA. Bovine pancreatic deoxyribonuclease I (DNase I) is a DNA minor grove-interacting nuclease, which shows relatively low specificity. This protein is composed of two central β sheets, each composed of six β-strands. This structure is surrounded by extensive loop and α-helical regions. This enzyme shares structural similarity to exonuclease III. DNase I is one of the most well characterized endonucleases of mammalian origin.

Biochem/physiol Actions

During tissue dissociation, parts of the cells are lysed resulting in a release of DNA. Monomolecular DNA may cause clumping of cells. Addition of DNase I to the dissociation buffer leads to a degradation of this extracellular DNA, therby avoiding the loss of cells from undesired clumping.

Unit Definition

一个单位表示在测定条件下导致吸光度以0.001每分钟的速率而增加的酶活力。

Preparation Note

激活剂：二价金属离子 (Ca²⁺, Mg²⁺)
工作浓度：0.01 至 1 mg/ml
注意：必须针对每种细胞类型单独确定工作浓度。为实现最佳活性，酶须添加 5 mM Mg²⁺。

Reconstitution

用无菌的双蒸水对冻干酶进行重悬（10 mg/ml）；用PBS（磷酸盐缓冲液）、HBSS（Hank′s 平衡盐溶液）或培养基进行进一步的稀释。
 

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主要经营的试剂品牌：
Roche试剂，部分常用现货，周三定货，2-3周到货。
GE试剂，耗材，部分常用现货，周二，周五定货，3周左右。
SIGMA试剂，周一，五定货，3周左右到货。
THERMO试剂，部分现货，周三定货，2-3周到货。
ABCAM试剂，国内现货三天左右，期货四周左右。
Peprotech试剂，部分常用现货，周三定货，2-3周到货。

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OXOID，BD培养基货期八周左右，部分现货。
MCE激动剂，ACROS、ALFA、阿拉丁等生化试剂现货定购。