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應用
胶原酶已用于动脉组织的制备,用于晚期糖基化终产物 (AGE) 的研究。这种酶还与其他蛋白酶一起用于人类肿瘤、小鼠肾、人脑、肺上皮和许多其他组织的解聚。在肝脏和肾脏灌注研究、胰腺的消化和非实质肝细胞的分离中也有效。也适用于大鼠附睾脂肪垫脂肪细胞的制备。
生化/生理作用
有效地从组织中释放细胞需要胶原酶和中性蛋白酶的共同作用。每摩尔酶用 4 克原子钙 (Ca2+) 激活胶原酶。一般认为培养滤液至少含有 7 种不同的蛋白酶,分子量范围为 68-130 kDa(最适用 pH:6.3-8.8)。该酶识别序列-R-Pro-8-X-Gly-Pro-R-,其中 X 是最常见的中性氨基酸。
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。
注意
供应的产品可在-20°C 下可稳定保存一年。在 37°C、50°C 和-20°C 条件下储存 30 天,FALGPA 或蛋白酶活性无损失。快速冷冻 (10 mg/mL) 并在-20°C 下冷冻时,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。
單位定義
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。
準備報告
本品由溶组织梭菌的 Ⅱ 型 (C6885) 胶原酶制备而成。通常在 37°C 下用 TESCA 缓冲液(含 50 mM TES,0.36 mM 氯化钙,pH 7.4)制备 1-2 mg/mL 溶液。溶液还含有梭菌蛋白酶、非特异性中性蛋白酶和胰蛋白酶活性。
基底
訊號詞
Danger
危險分類
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
標靶器官
Respiratory system
儲存類別代碼
11 - Combustible Solids
水污染物質分類(WGK)
WGK 1
閃點(°F)
Not applicable
閃點(°C)
Not applicable
個人防護裝備
dust mask type N95 (US), Eyeshields, Faceshields, Gloves
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实验方案
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.
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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