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产品说明/Specification Sheet
产品名称: |
Dispase II |
产品编号: |
04942078001-500mg |
规格: |
500mg |
价格: |
¥750 |
产品类别: |
生化试剂 |
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品牌: |
ROCHE |
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产品属性:
Dispase即为中性蛋白酶,也称分散酶,是一种非特异性的金属蛋白酶,用于从不同组织或器官中消化细胞外基质,释放并制备原代单细胞;或用于细胞培养中的细胞收获或接种转移;也被用于防止悬浮细胞培养时的细胞意外结团。
常用的胰蛋白酶、胶原酶和链酶蛋白酶都属于蛋白水解酶类,用于分散组织、分离细胞,但其对细胞伤害大,细胞在培养时不稳定,甚至引入支原体污染。Dispase不会伤害细胞膜,且因为其来源于细菌,故不会引入支原体或任何动物病毒;温度、pH、血清成分等的差异对Dispase影响很小。Dispase经稀释后其活性就大大降低,这也方便了后续的培养。
Dispase优点:
该酶作用迅速有效又温和;
细胞能保持完整,无损细胞膜;
该酶为细菌来源,不含支原体或动物病毒污染;
对于温度、pH的改变、以及血清成分都很稳定;
使用螯合剂或稀释即可简单灭活。
酶活定义:
1U是在检测条件下1分钟释放相当于1 umol酪氨酸的Folin阳性氨基酸或肽的酶量;DispaseII:≥0.8 U/mg (37度,使用酪蛋白作为底物,pH 7.5);
抑制剂:EDTA、EGTA、Hg2+、其他重金属。血清不会抑制酶活性。
使用方法:
用Hepes-buffered saline (50 mM Hepes/KOH pH 7.4, 150 mM NaCl)溶解Dispase至10 mg/ml,使用时用细胞培养液稀释到0.6-2.4 U/ml。不推荐高于2.4 U/ml的工作浓度,可用0.22 um的无菌滤膜过滤除菌。
保存方法:
配成溶液后,4度保存,2周内使用有效,-20度保存,2个月内使用有效,不要反复冻融;储存液用Hepes盐缓冲液稀释后,4度保存,3天内使用有效。
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2012/6/5 9:40:36 (蓝博斯特-丁) | 110.96.167.* | 详细信息:
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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