溶组织梭菌来源中性蛋白酶属于蛋白水解酶，可通过组织解离实现多种细胞类型的分离。
高活性级中性蛋白酶 NB为经色谱纯化的金属蛋白酶，适用于各类组织解离实验。本品蛋白水解活性高，基本无胶原酶活性（PZ 单位检测），内毒素水平极低。用于细胞分离时，可与高纯度胶原酶 NB 1 优品级、胶原酶 NB 4 标准级或胶原酶 NB 4G 证明等级搭配使用。
组分：含有经色谱纯化的高活性中性蛋白酶。
应用
高活性级中性蛋白酶 NB 通常搭配胶原酶 NB 1 优品级 （货号 C278986）或其他胶原酶 NB 系列产品使用，适用于从各类组织中分离细胞。
使用说明
概述
高活性级中性蛋白酶 NB，搭配胶原酶 NB 1 优品级使用，特别适用于从人胰腺中分离朗格汉斯胰岛，也可用于多种其他人体或动物组织的细胞分离。
组织解离
从人胰腺分离朗格汉斯胰岛的推荐起始浓度：
胶原酶 NB 1 优品级：15 – 20 PZ U /g 组织
高活性级中性蛋白酶 NB：0.8 – 1.5 DMC U /g 组织
一般来说，合适的中性蛋白酶浓度取决于组织类型、来源以及分离操作方案。
中性蛋白酶的活性在 37℃条件下达到最佳。
储备液（母液）
高活性级中性蛋白酶 NB 储备液配制：取适量酶溶于合适体积的纯水。 酶溶液需全程置于冰上保存。
该中性蛋白酶活性依赖钙离子，体系中严禁存在任何钙螯合剂（例如 EDTA）。
复溶后的高活性中性蛋白酶可过滤、分装，在 -20℃条件下最长可保存 1 年；应避免反复冻融。
推荐使用低蛋白结合特性的 0.22 μm 滤膜（如醋酸纤维素、PVDF 或 PES 滤膜）进行过滤。
工作液
配制工作液：用水稀释储备液至所需中性蛋白酶浓度。工作液使用前需全程冰上存放。
若将高活性级中性蛋白酶 NB 溶液与胶原酶 NB 溶液混合，混合液需立即使用。
失活与抑制剂
可通过降温或者稀释酶溶液的方式减弱解离反应。 中性蛋白酶抑制剂包含 EDTA 等螯合剂。
重要提示
高活性级中性蛋白酶 NB 仅用于科研。
General Neutral protease from Clostridium histolyticum is a proteolytic enzyme and therefore can be used for isolation of various cell types by tissue dissociation.
Neutral Protease NB High Active Grade is a chromatographically purified metalloprotease suitable for different tissue dissociation applications. This product contains an increased proteolytic activity, is largely free from collagenolytic activity (PZ U) and has a very low endotoxin level. For cell isolation, it is used in combination with highly purified Collagenase NB 1 Premium Grade, Collagenase NB 4 Standard Grade or Collagenase NB 4G Proved Grade.
Composition: Contains highly active, chromatographically purified neutral protease.
Application
Neutral Protease NB High Active Grade is, mostly in combination with Collagenase NB 1 Premium Grade (Cat. No. C278986) or another Collagenase NB product, suitable for cell isolation from various tissue types.
Instructions for use
General
Neutral Protease NB High Active Grade is, in combination with Collagenase NB 1 Premium Grade, especially suited for isolation of islets of Langerhans from human pancreas, and also for cell isolation from several other human or animal tissues.
Tissue dissociation
Recommended starting concentrations for isolation of islets of Langerhans from human pancreas:
Collagenase NB 1 Premium Grade: 15 – 20 PZ U/g tissue
Neutral Protease NB High Active Grade: 0.8 – 1.5 DMC U/g tissue
In general, the appropriate neutral protease concentration depends on tissue type and origin as well as on the isolation procedure.
Neutral protease activity is at an optimum at 37 °C.
Stock solution
A stock solution of Neutral Protease NB High Active Grade is prepared by dissolving the enzyme in an appropriate volume of water. The enzyme solution must be constantly stored on ice.
Since neutral protease depends on calcium, absolutely no calcium chelating agents (e.g. EDTA) should be present at all.
Reconstituted Neutral Protease High Active Grade can be filtered, aliquoted and stored at -20 °C for up to 1 year.
Repeated freezing and thawing should be avoided. For 0.22 µm filtration filters with low protein binding properties (e.g. cellulose acetate, PVDF, or PES) are recommended.
Working solution
To prepare a working solution, the stock solution is diluted with water to achieve the required neutral protease concentration. The working solution must be constantly stored on ice until use.
If Neutral Protease NB High Active Grade solution is mixed with Collagenase NB solution, the blend should be used immediately.
Inactivation and inhibitors
The dissociation process can be reduced, e.g. by cooling down or dilution of the enzyme solution. Inhibitors of neutral protease include chelating agents like EDTA.
Important
Neutral Protease NB High Active Grade is intended for research use on
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