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In stock Molecular Biology Reagents 翌圣生物 Yeasen

Collagenase I 胶原酶I型

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酶消化法原代细胞制备细胞解离

Product no.40507ES Sizes2 options SourceYeasen · Official

Cat. No. 40507ES60

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40507ES60 100 mg ¥335.00促~~¥365.00~~ 现货 Enquire

40507ES76 1 g ¥3,285.00 现货 Enquire

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Overview

Specifications

Brand翌圣生物 Yeasen

Cat.No40507ES

Specification酶活力单位定义在37℃，pH7.5 的条件下，5 h内水解胶原产生相当于1 µM L-亮氨酸的酶量定义为1个酶活力单位。

Origin翌圣生物科技(上海)股份有限公司

技术平台 酶消化法

应用方法 原代细胞制备用于上皮、肝、肺、脂肪和肾上腺组织来源细胞的分离和制备 细胞解离降解胶原纤维，解离组织细胞外基质

生物特性参数 细胞产品子类细胞分离与消化酶 细胞类型上皮细胞、肝、肺、脂肪和肾上腺组织细胞 应用用于组织和细胞的解离，尤其适用于上皮、肝、肺、脂肪和肾上腺

样本类型 组织细胞

技术参数 酶活力≥250 U/mg solid 来源溶组织梭菌 产品类型胶原酶I型 储存条件2~8℃避光保存，2年有效；储存液-20℃避光冻存 运输条件室温运输

实验类型 组织分离利用胶原酶I型消化组织块，制备单细胞悬液 器官灌注通过灌注胶原酶工作液解离器官细胞 皮肤单细胞悬液制备与分散酶和DNase联合消化皮肤组织，获得单细胞

FAQ

Q：只用于分离细胞，选哪种胶原酶合适？

A：对于组织解离，常使用粗制胶原酶制剂，例如弹性蛋白酶，胰蛋白酶或木瓜蛋白酶等；对于胶原蛋白结构和生物合成研究，常使用更高纯度的胶原酶制剂，不含其他蛋白水解活性。

Q：要把肿瘤细胞分散成单个细胞做流式检测，选择哪一种胶原酶比较适合？需要区分肿瘤发生的部位吗？后续用来做流式，不同消化强度对细胞表面抗原的影响大吗？

A：会影响细胞表面抗原，可以选择对抗原影响较小的胶原酶Ⅲ、胶原酶Ⅳ。

Q：胶原酶使用会受到血清影响吗？

A：胶原不会受到培养细胞所用的血清影响。

Q：胶原酶的使用会受哪些抑制剂影响？

A：胶原酶的抑制剂有 EDTA 、EGTA 等；二是重金属离子（Hg²⁺,Pb²⁺,Cd²⁺）；还有一些化学试剂如 Cysteine、 histidine、DTT、2-mercaptoethanol、o-phenanthroline 等对胶原酶也有抑制作用。

Q：用 PBS 溶解可以吗？用什么工作液浓度溶解？

A：建议在 Hank’s，Earle’s 或者其他平衡盐溶液中溶解，推荐使用工作浓度 0.05 % ~0.5 % (w/v)。

Q：实验过程中消化作用不佳

A：

(1)如果是酶活不强或钙离子不足导致，应在低温干燥条件下储存胶原酶，胶原酶溶液分装后冻存

(2)如果是钙离子不足导致，应将胶原酶溶液中钙离子浓度设置为 5 mM。

Q：使用过程中出现细胞死亡

A：

(1)如果是蛋白酶过量，应减少蛋白酶接触时间或是加入白蛋白或灭活血清；

(2)如果是pH改变应使用缓冲液（如 HBSS）；经常检查并重新调节 pH；

(3)如果氧气太少，应给消化液通无菌空气。

Q：实验中出现细胞受损怎么办？

A：如果是蛋白酶过多导致，应减少蛋白酶用量或是加入白蛋白或灭活血清

Documents & Citations

Published citations

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