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Merck
CN

C0773

胶原酶 来源于溶组织梭菌

high purity, purified by chromatography, Type VII, ≥4 FALGPA units/mg solid, lyophilized powder, ≥700 CDU/mg solid (CDU = collagen digestion units)

别名:

梭菌肽酶 A

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关于此项目

化学文摘社编号:
UNSPSC Code:
12352204
NACRES:
NA.54
EC Number:
232-582-9
MDL number:
Specific activity:
≥4 FALGPA units/mg solid, ≥700 CDU/mg solid (CDU = collagen digestion units)
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Quality Level

type

Type VII

form

lyophilized powder

specific activity

≥4 FALGPA units/mg solid, ≥700 CDU/mg solid (CDU = collagen digestion units)

mol wt

68-130 kDa

purified by

chromatography

composition

Protein, ≥85% biuret

solubility

TESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4): soluble 0.05-0.1 mg/mL at 37 °C

foreign activity

caseinase and clostripain ≤1 units/mg protein

storage temp.

−20°C

Application

胶原酶已用于制备研究晚期糖基化终产物(AGE)的动脉组织。该酶还与其他蛋白酶一起用于人肿瘤、小鼠肾、人脑、肺上皮和许多其他组织的解聚。它在肝脏和肾脏灌注研究、胰腺消化和非实质肝细胞的分离中也是有效的。

Biochem/physiol Actions

从组织中有效释放细胞需要胶原酶和中性蛋白酶的共同作用。胶原酶被4克原子钙(Ca2+)/摩尔的酶活化。培养滤液至少含有7种分子量为68-130kDa的不同蛋白酶。最佳pH范围为6.3-8.8。该酶识别序列 -R-Pro-8-X-Gly-Pro-R-,其中X通常是中性氨基酸。
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。

Physical form

含氯化钙的冻干粉

Preparation Note

储液制备:将0.05-0.1 mg胶原酶溶于1 mL含0.36 mM氯化钙的50 mM TES缓冲液(TESCA)中,pH 7.4(37 °C)。采用冷的缓冲液。
请注意,材料不会完全溶解,会出现少量沉淀。沉淀并不会影响溶液的活性。

Other Notes

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。


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pictograms

Health hazardExclamation mark

signalword

Danger

Hazard Classifications

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

target_organs

Respiratory system

存储类别

11 - Combustible Solids

wgk

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

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

法规信息

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分析证书(COA)

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实验方案

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。

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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