Dispase® II (neutral protease, grade II) 分散酶（中性蛋白酶）
应用
蛋白水解酶，可用于从广泛组织中分离制备细胞。该酶可快速、有效、温和地去除组织中的细胞外结构，释放出单个细胞，适用于组织细胞的剥离及将收获的细胞转种到新的培养基中做后续的培养。另外，还可用于避免细胞悬浮培养中不必要的细胞聚集。
Dispase is used for the preparation of cells from a wide variety of different tissues and organs. The enzyme has proven to be a rapid and effective, yet gentle agent for the disruption of extracellular matrix of tissues for releasing single cells for cell culture (primary cell culture) or harvesting cells already in culture to transfer to new substrate (secondary culture). Furthermore, it is used to prevent unwanted clumping of cells cultured in suspension.
优点
Rapid, effective, yet gentle agent that liberates cells with minimal cell damage
Maintains cell membrane integrity
Non-mammalian source - free of mycoplasma and animal virus contamination
Extremely stable to influences of temperature, pH, and interference by serum components
Easily inactivated by chelating agents or by dilution
Delivers higher activity and convenience
产品描述
EC 3.4.24.4
Enzyme activity: ≥0.8 U/mg (37°C, casein as substrate, pH 7.5)
Inhibitors: EDTA, EGTA, Hg2+, other heavy metals. Dispase is not inhibited by serum.
Specificity: Dispase is a nonspecific metalloprotease.
Working concentration: 0.6 to 2.4 U/ml
背景信息
One unit is the enzyme activity which liberates Folin-positive amino acids and peptides corresponding to 1 µmol tyrosine in 1 minute under assay conditions. A practical comparison of Roche units of Dispase® II with those cited in the Japanese literature (where dispase concentrations of 1,000 to 2,000 units/ml are not uncommon) suggests that one Roche unit equals approximately 600 Japanese units of dispase.
工作浓度
Dissolve the non-sterile lyophilized enzyme in Hepes-buffered saline (50 mM Hepes/KOH pH 7.4, 150 mM NaCl) (10 mg/ml). Dilute further with the culture medium to be used for the isolated cells to a final concentration of 0.6 to 2.4 U/ml. Concentrations higher than 2.4 U/ml are not recommended. Sterilize through a 0.22 μm filter membrane.
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