Collagenase is a protease that cleaves the bond between a neutral amino acid (X) and glycine in the sequence Pro-X-Gly-Pro, which is found with high frequency in collagen. Collagenase is unique among proteases in its ability to degrade the triple-helical native collagen fibrils commonly found in connective tissues such as skin, tendon, blood vessels, and bone. Collagenase disaggregation is suitable for the culture of human tumors, mouse kidney, human adult and fetal brain, and many other tissues including epithelium. Collagenase is relatively gentle, dissociates well at physiological temperature and pH, and requires no mechanical agitation or special equipment.
Collagenase Type I is isolated from Clostridium histolyticum and packaged as a lyophilized, non-sterile powder for research use in cell or tissue dissociation, and organ perfusions. Collagenase Type I activity is guaranteed to be greater than 125 units/mg. Compared to other collagenase preparations, Collagenase Type I has average levels of collagenase, caseinase, clostripain, and tryptic activities, and is well-suited for the digestion of fat, adrenal, and liver cells or tissues.
Reagent Type: Collagenase
Shipping Condition: Room Temperature
Storage conditions: 2°C to 8°C. Protect from light.
Shipping conditions: Room temperature
Shelf life: 24 months from date of manufacture
产品介绍
本公司胶原酶来源于进口原料,酶活性高,纯度高。本品为固体粉末状态。
胶原酶Ⅰ,用于上皮、肺和肾上腺组织细胞的分离。外观为黑棕色粉末。
活性: 高于125 units/mg
分子量68,000 到125,000,
最适PH为6.3-8.8。
胶原酶能够特异性结合-R-Pro-8-X-Gly-Pro-R-序列中的中性氨基酸(X) 和甘氨酸之间的肽键。该粗品是溶组织梭菌分泌的混合酶。
主要为胶原酶、中性蛋白酶和梭菌蛋白 酶。其中梭菌蛋白酶是被氧化的、最不活泼的酶,组织的分离必须依靠胶原酶和中性蛋白酶两者共 同作用。
使用说明:
胶原酶Ⅰ可以用 TESCA buffer (50 mM TES,0.36 mM 氯化钙,pH 7.4,37°C)配制成 1-2 mg/ml。
也可以用含钙镁的 PBS、含钙镁的 hanks 或不含血清的培养基配制。(配完后分装,避免反复冻融)。
活性说明:
1 个胶原消化单位(CDU):在 pH 7.4,37℃以及钙离子存在的条件下,以每5小时从胶原中 释放的多肽相当于茚三酮显色1微摩尔亮氨酸量为一个活性单位。
1FALGPA 单位:在 pH 7.5、25℃以及钙离子存在的条件下,一个 FALGPA 水解单位每分钟 可水解 1.0μmol 呋喃基丙烯酰基-Leu-Gly-Pro-Ala。
激活剂:Ca2+,最适合浓度为5 mM。
抑制剂:EGTA、还原性谷胱甘肽、β-巯基乙醇、巯基乙酸钠、8-羟基喹啉、2,2'-联吡啶等。
底物:不同类型的胶原是其天然底物,而一些合成的多肽也可以作为其底物,比如: N-CBZ-gly-pro-gly-gly-pro-ala (Km = 0.71 mM)、N-CBZ-gly-pro-leu-gly-pro、N-(3-(2-furyl)acryloyl)- leu-gly-pro-ala(FALGPA)、4-Phenylazo benzyloxycarbonyl-pro-leu-gly-pro-D-arg、N-2,4-Dinitrophenyl- pro-gln-gly-ile-ala-gly-gln-D-arg。除此之外N-Succinyl-gly-pro-leu-glypro7-amido-4-methylcoumarin是 类胶原酶的肽酶的底物。N-(2,4-Dinitrophenyl)-proleu-gly-leu-trp-ala-D-arg 氨基酸化合物是脊椎动物 胶原酶的底物。