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Merck

D4693

Sigma-Aldrich

分散酶®II

protease

别名:

蛋白酶 来源于多粘芽孢杆菌

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About This Item

CAS号:
EC號碼:
MDL號碼:
分類程式碼代碼:
12352204
NACRES:
NA.54

品質等級

形狀

lyophilized solid

比活性

≥0.5 units/mg solid

技術

cell culture | mammalian: suitable
single cell analysis: suitable

溶解度

10 mM NaAc (pH 7.5) and 5 mM CaAc: soluble

儲存溫度

2-8°C

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應用

来自Sigma的该酶已被用于开发用于离体培养小鼠胚胎乳腺芽的实验方法。它已被用于在从大鼠心脏分离线粒体的过程中对大鼠心脏碎片进行处理。它还被用于通过酶水解来分离牙髓干细胞(DPSC)。这些细胞已与通过外植体方法而分离的DPSC进行了比较,以分析它们的干细胞和分化特性。
中性蛋白酶® II已用于荧光激活细胞分选(FACS)。它还被用于分离内脏卵黄囊层。
适用于制备测序用单细胞悬浮液。

生化/生理作用

中性蛋白酶®II是一种中性的蛋白酶,可水解非极性氨基酸残基的N-末端肽键。它可用于分离生长在体外的多种组织和细胞。该酶非常温和,不会对细胞膜造成损伤。它还可用于防止悬浮培养物中的细胞结块。该蛋白酶可水解纤连蛋白和IV型胶原蛋白,但不能水解层粘连蛋白、V型胶原蛋白、血清白蛋白或转铁蛋白。中性蛋白酶®II具有切割亮氨酸-苯丙氨酸键的特异性。Ca2+、Mg2+、Mn2+、Fe2+、Fe3+和Al3+ 可灭活分散酶I。EDTA、EGTA、Hg2+ 和其他重金属可抑制该酶活性。每g-mol纯化分散酶I含有1g锌原子。如果通过EDTA或EGTA等螯合剂除去锌成分,则产生无活性的酶蛋白。该酶不受血清抑制。

單位定義

除特殊说明,在pH 7.5,37 °C 条件下,一单位的酶每分钟将会水解酪蛋白以产生与1.0 μ摩尔 (181 μg) 酪氨酸等同的颜色(Folin-Ciocalteu试剂提供的颜色)。

外觀

含有乙酸钙和乳糖的冻干粉

準備報告

可通过将粉末溶解在含有10 mM NaAc (pH 7.5)和5 mM CaAc的缓冲液中制备50 U/mL的储备溶液。它应当被储存在4 °C。

分析報告

该酶初始可以10 mg/ml溶解于50 mM Hepes/KOH pH 7.4, 150 mM NaCl中。 进一步的稀释应当在最适于待解离细胞的培养基中进行。 中性蛋白酶是一种金属蛋白酶,可通过包括Ca2+ , Mg2+, Mn2+以及Fe2+在内的二价阳离子激活。

法律資訊

Dispase is a registered trademark of Godo Shusei Co., Ltd.

象形圖

Exclamation markHealth hazard

訊號詞

Danger

危險分類

Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3

標靶器官

Respiratory system

儲存類別代碼

11 - Combustible Solids

水污染物質分類(WGK)

WGK 3

閃點(°F)

Not applicable

閃點(°C)

Not applicable


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