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安全信息

C0130

Sigma-Aldrich

胶原酶 来源于溶组织梭菌

for general use, Type I, ≥125 CDU/mg solid

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别名:
梭菌肽酶 A
CAS号:
9001-12-1
EC 号:
3.4.24.3 (BRENDA, IUBMB)
EC 号:
232-582-9
MDL编号:
MFCD00130830
NACRES:
NA.54

形式

lyophilized powder

specific activity

≥125 CDU/mg solid

分子量

68,000 to 125,000

最佳pH

6.3-8.8

溶解性

TESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4): soluble 1-2 mg/mL at 37 °C

储存温度

2-8°C

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相关类别

一般描述

胶原酶是一种可切割被称为胶原蛋白的三螺旋蛋白的蛋白酶。存在有三种类型的组织胶原酶，而它们都属于基质金属蛋白酶（MMP）家族。从溶组织梭菌中获得的胶原酶具有非常强的活性，因为它可在低至4-10℃的温度下从两端对胶原蛋白进行消化。粗胶原酶混合物含有两种主要的酶类型，即胶原酶和梭菌蛋白酶。

应用

来自溶组织梭菌的胶原酶在生物学研究中具有广泛的用途，它可用于制备分离的细胞悬浮液。该产品适用于人类肿瘤、小鼠肾脏、人类成人和胎儿脑、肺和许多其他上皮组织的解聚。它还被证明可有效用于肝脏和肾脏灌注研究、胰腺消化、非实质大鼠肝细胞的分离以及肝细胞的制备。 胶原酶已用于研究晚期糖基化终产物所需的动脉组织的制备。该酶已经测试可以约1 mg/mL的浓度释放肝细胞。用于消化的浓度范围是0.1至5 mg/mL。

生化/生理作用

该胶原酶产物是由溶组织梭菌所分泌的酶混合物组成，且不同的产物可通过在分泌酶中所发现的10-18种化合物的比例而进行区分。 主要的成分是两种胶原酶、梭菌蛋白酶和一种中性蛋白酶。 这些酶的协同作用可降解胶原蛋白及其他细胞内物质。胶原酶和中性蛋白酶的作用对于从组织中有效释放细胞是必需的。 不同类型的胶原蛋白是胶原酶的天然底物。
每摩尔胶原蛋白酶被4g原子钙活化。 它被乙二醇-双（β-氨基乙基醚）-N，NN′,N′-四乙酸，β-巯基乙醇、谷胱甘肽、巯基乙酸和8-羟基喹啉抑制。

注意

根据所提供的信息，该产品在-20°C下可稳定保存一年。 在37℃、50℃和-20℃下FALGPA或蛋白酶的活性在30天内不会有损失。如果进行分装（10mg / mL）并快速冻存在-20℃，则粗胶原酶的溶液是可以稳定的。进一步的反复冻融会损坏溶液。 该产品在置于冰上时可保持100%活性超过7小时。

单位定义

一个胶原蛋白消化单元（CDU）从牛跟腱胶原蛋白中释放的肽链量和1.0 μ亮氨酸，5小时，pH7.4,37℃，在钙离子存在下的茚三酮颜色相同。一个 FALGPA水解单元每分钟，25℃，水解1.0 μmole 呋喃基丙烯酰-Leu-Gly-Pro-Ala。一个中性蛋白酶单元水解酪蛋白，产生颜色和1.0 μmole 酪氨酸每5小时，pH7.5,37℃下产生的相同。一个梭菌蛋白酶单元在DTT存在下，pH7.6,25℃，每分钟水解1.0 μmole BAEE。

制备说明

该产品相当于最初的40％硫酸铵部分 Mandl, I., et al.,J. Clin.Invest., 32, 1323 (1953).溶液通常是在TESCA缓冲液（含有50mM TES、0.36mM氯化钙，37℃时pH 7.4）中按1-2 mg/mL进行制备。

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象形图

Health hazard
GHS08

警示用语：

Danger

危险声明

H334

预防措施声明

P261 - P284 - P501

危险分类

Resp. Sens. 1

储存分类代码

11 - Combustible Solids

WGK

WGK 1

闪点(°F)

Not applicable

闪点(°C)

Not applicable

个人防护装备

dust mask type N95 (US), Eyeshields, Faceshields, Gloves

法规信息

常规特殊物品

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  1. How should I prepare a stock solution of Product C0130, Collagenase?

    Stock solutions of crude Collagenase may be prepared at 10 mg/ml of Collagenase in TESCA buffer (50 mM TES, 0.36 mM calcium chloride, pH 7.4 at 37°C) and are stable if frozen quickly in aliquots and kept frozen at -20°C. Freeze-thaw cycles will damage the enzyme.

  2. How should Product C0130, Collagenase, be prepared for use in tissue digestions?

    For use in tissue culture applications, Collagenase can be solubilized in calcium-free solutions such as Hanks Balanced Salts (H2387) or Earl's' Balanced Salt Solution (E6267).

  3. What is the activity of the lot number of Product C0130, Collagenase, which I have?

    The activity and specific activity for each lot number can be obtained from the certificate of analysis.

  4. What type of Collagenases are present in Product C0130, Collagenase,?

    This Collagenase is obtained from the culture filtrate of Clostridium histolyticum. The culture filtrate is thought to contain at least 7 different proteases ranging in molecular weight from 68 - 130 kDa. It does not contain any Matrix Metalloprotease.

  5. Is Product C0130, Collagenase, sterile?

    This product does not consist of a sterile preparation of Collagenase. Upon reconstitution, solutions should be sterile filtered through a 0.22 micron filter.

  6. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  7. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

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    Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product. 

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实验方案

使用FALGPA（N-（3- [2-呋喃基]丙烯酰基）-Leu-Gly-Pro-Ala）进行的胶原酶酶法检测（EC 3.4.24.3）

在测定胶原酶活性时，使用N-（3-[2-呋喃基]丙烯酰基）-Leu-Gly-Pro-Ala在345nm处进行连续分光光度法测定。胶原酶水解胶原肽键。

Enzymatic Assay of Collagenase (EC 3.4.24.3) using FALGPA (N-(3-[2-Furyl]acryloyl)-Leu-Gly-Pro-Ala)

To measure collagenase activity, N-(3-[2-Furyl]acryloyl)-Leu-Gly-Pro-Ala is used in a continuous spectrophotometric rate determination at 345 nm. Collagenase hydrolyzes collagen peptide bonds.

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