来源于溶组织梭菌的胶原酶属于蛋白水解酶，可原位切割人或动物组织中三螺旋胶原分子的肽键。
因此，胶原酶被广泛应用，通过组织解离分离多种类型细胞。
胶原酶 NB 4G（证明等级）属于粗制胶原酶，具备胶原水解活性，同时含有梭菌蛋白酶、中性蛋白酶等次级酶活性；各类酶活性配比均衡，可实现温和且高效的组织解离。
应用
胶原酶 NB 4G（证明等级）适用于多种组织类型的解离。
使用说明
1、概述：
胶原酶 NB 4G（证明等级）适用于从人或动物组织中分离多种细胞；适用组织包括皮肤、肝脏、结肠肿瘤、非洲爪蟾卵巢。
2、组织解离：
部分应用推荐起始工作浓度：人 / 啮齿类皮肤：0.2‑0.5 PZ U/mL；啮齿类肝脏：0.1‑0.15 PZ U/mL。
一般而言，合适的胶原酶浓度取决于组织类型、组织来源以及分离方案。
胶原酶最适反应条件：37 ℃，pH 7.4。
3、母液配制：
在细胞分离常用缓冲液中，本品最高可配至 150 mg/mL；酶溶液全程置于冰上保存。
胶原酶及部分次级蛋白酶活性依赖钙离子，建议缓冲液钙离子浓度≥2 mM；体系严禁存在钙离子螯合剂（例如 EDTA）。
如需 0.22 μm 过滤除菌，推荐选用低蛋白吸附滤膜（醋酸纤维素 CA、聚偏二氟乙烯 PVDF、聚醚砜 PES）。
4、工作液：
取母液用缓冲液稀释至目标胶原酶活性浓度；工作液使用前全程冰上放置。
5、酶灭活与抑制剂：
可通过降温或者稀释酶液减缓解离反应。高 pH 条件下胶原酶可逆失活；低 pH 条件下发生不可逆失活。胶原酶抑制剂包括半胱氨酸、EDTA 等螯合剂。
6、重要提示：
胶原酶 NB 4G（证明等级）仅用于科学研究。
Collagenases from Clostridium histolyticum are proteolytic enzymes that cleave peptide bonds in the triple helical collagen molecule of human or animal tissue in‑situ.
For this reason collagenases are widely used for isolation of various cell types by tissue dissociation.
Collagenase NB 4G Proved Grade is a crude collagenase which contains collagenolytic and additional enzymatic activities including clostripain and neutral protease. The balanced ratio of these activities leads to gentle and efficient tissue dissociation.
Application
Collagenase NB 4G Proved Grade is suitable for dissociation of a broad variety of tissue types.
Instructions for use
1. General:
Collagenase NB 4G Proved Grade is suitable for isolation of a broad variety of cells from human or animal tissues. Tissue types include skin, liver, colon tumors, and xenopus ovaries.
2. Tissue dissociation:
Recommended starting concentrations for selected applications:
Skin (human, rodent): 0.2 – 0.5 PZ U/ml；Liver (rodent): 0.1 – 0.15 PZ U/ml
In general, the appropriate collagenase concentration depends on tissue type and origin as well as on the isolation procedure.
Collagenase activity is at an optimum at 37 °C and pH 7.4.
3. Stock solution:
Collagenase NB 4G Proved Grade dissolves at a concentration of up to 150 mg/ml in all buffers which are commonly used for cell isolation. The enzyme solution must be constantly stored on ice.Since collagenase and some of the secondary proteases depend on calcium, it is recommended to use a buffer with ≥ 2 mM Ca²⁺. Absolutely no calcium chelating agents (e.g. EDTA) should be present at all.For 0.22 µm filtration filters with low protein‑binding properties (e.g. cellulose acetate, PVDF, or PES) are recommended.
4. Working solution:
To prepare a working solution, the stock solution is diluted with buffer to achieve the required collagenase concentration. The working solution must be stored on ice until use.
5. Inactivation and inhibitors:
The dissociation process can be reduced, e.g. by cooling down or dilution of the enzyme solution. Collagenase is reversibly inactivated at high pH values and irreversibly inactivated at low pH values. Inhibitors of collagenase include cysteine or chelating agents like EDTA.
6. Important note:
Collagenase NB 4G Proved Grade is intended for research use only.
K2306413
危险品化学品经营许可证（带存储）