应用
该产品适用于人类肿瘤,小鼠肾脏,人类成人和胎儿的脑、肺和许多其他上皮组织的解聚。 它还被证明适用于肝脏和肾脏灌注研究、胰腺消化、非实质性大鼠肝细胞的分离、以及肝细胞制备。 胶原酶也已用于制备动脉组织,以对晚期糖基化终产物进行研究。 该酶用于(浓度约为1mg/ml)对肝细胞的释放进行检测。 消化浓度范围为0.1至5 mg/mL。
适用于制备用于测序的单细胞悬液。
适用于制备用于测序的单细胞悬液。
生化/生理作用
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。
胶原酶产品是由溶组织梭菌分泌的一种混合酶 ,根据分泌的酶中 10-18 组分的相对比例而区分不同的产品。主要成分为两种胶原酶— 梭菌蛋白酶和一种中性蛋白酶。这些酶的协同操作降解胶原和胞内其他物质。胶原酶和中性蛋白酶的作用是使细胞有效地从组织中释放出来所必需的。各种类型的胶原是胶原酶的天然底物。
注意
供应时,本品在-20℃ 下可稳定保存一年。在 37°C、50°C 和-20°C 条件下储存 30 天后,FALGPA 或蛋白酶活性未损失。如果快速冷冻 (10 mg/mL) 并在-20°C 下冷冻,粗胶原酶溶液是稳定的。进一步冻融循环将破坏溶液。置于冰上 7 小时后,产品仍保持 100% 活性。
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
通常在 37°C 下用含 50 mM TES、0.36 mM 氯化钙 (pH 值为 7.4) 的 TESCA 缓冲液制备 1-2 mg/mL 溶液。本品还含有梭菌蛋白酶、非特异性中性蛋白酶和胰蛋白酶活性。
警示用语:
Danger
危险分类
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
靶器官
Respiratory system
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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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。
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