计算溶液所需的质量、体积或浓度。
概述
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阿拉丁和生产的EnzymoPure™ Express Enzyme，即EnzymoPure™ Express Enzyme，是一种使用室温下稳定的重组类胰蛋白酶(Recombinant trypsin-like enzyme, 简称TrypsinLE或TrypLE)并且不含动物源性物质(Animal origin-free, AOF)的即用型细胞消化液(Cell dissociation solution)。本产品消化温和、快速有效、对细胞几乎无损，可以完美替代普通胰酶细胞消化液。
本产品经过过滤除菌处理，含约1mM EDTA，pH值为7.2-7.8，不仅可以和普通的胰酶细胞消化液一样直接用于培养细胞或一些组织的消化，更适合用于一些严格要求使用无动物源性胰酶消化液的细胞或组织的消化。
胰蛋白酶是一种丝氨酸蛋白酶，可特异切割赖氨酸及精氨酸C末端肽键。阿拉丁的EnzymoPure™ Express Enzyme使用的是高纯度的重组类胰蛋白酶，也称胰酶替代物(Trypsin replacement)，蛋白序列来源于真菌，具有与动物源性猪胰蛋白酶相同的酶学性质，且稳定性更高，纯度更好，消化更温和，细胞毒性更低，可替代猪胰腺来源胰蛋白酶应用于细胞和组织的消化等用途。
本产品使用的重组类胰蛋白酶无动物源性。重组生产，不含动物源性与人源成份，无动物源性的病毒污染，生产过程不使用任何动物源性原料。宿主蛋白残留量和DNA残留量都符合药典标准。传统胰酶大多取自猪、牛的胰脏，胰脏中除了胰蛋白酶，还含有多种其它的蛋白酶，而且传统胰酶生产工艺的特点决定了胰酶中不可避免地含有动物来源的其它杂质。
本产品的消化效果好。相同条件下，本产品中的重组类胰蛋白酶酶活性比普通的含0.05%胰酶的胰酶细胞消化液要高约2倍，对于多数细胞的消化效果也更好。通常室温1-3分钟左右就可以消化下大多数贴壁细胞。使用本产品消化细胞的效果参考图1。通常在细胞出现明显的轻微形态变化，并且细胞并未完全变圆时，即可吹打下细胞。
图1. 阿拉丁EnzymoPure™ Express Enzyme消化不同细胞的效果图。使用本产品分别消化A498、BEAS-2B、L929细胞。在1-2分钟内，显微镜下可观察到细胞边缘明显收缩，紧密贴合的细胞离散成单个细胞。注：本实验中，EnzymoPure™ Express Enzyme室温放置10分钟左右，未经预热；细胞未经PBS洗涤，在吸净细胞培养液后直接加入适量EnzymoPure™ Express Enzyme在约25℃的室温进行消化。如果用PBS或HBSS等洗涤细胞用于去除残余的血清，37℃消化效果会更好。实际效果会因细胞和实验条件的不同而略有差异，本图仅供参考。
本产品使用的重组类胰蛋白酶纯度高，对细胞作用温和，即使过度消化后对细胞损伤也非常小。通过亲和层析等多个纯化步骤获得的高纯度重组类胰蛋白酶，酶比活高，减少了猪胰蛋白酶提取物中存在的其它酶对细胞可能造成的损害，而且本产品无需使用胰酶抑制剂或血清对其进行终止。即使过度消化后对细胞损伤也非常小 (图2) 。
图2. 阿拉丁EnzymoPure™ Express Enzyme过度消化细胞后细胞的生长效果图。用EnzymoPure™ Express Enzyme分别消化A498、BEAS-2B细胞，过度消化(Over-digestion)细胞至全部漂浮，未使用胰蛋白酶抑制剂，离心后收集细胞沉淀，重新铺板后，继续培养18小时，显微镜下可观察到EnzymoPure™过度消化组和正常消化组无明显差异，贴壁较好。对照为正常消化的细胞(Normal-digestion)。实际效果会因细胞和实验条件的不同而略有差异，本图仅供参考。
本产品十分稳定。本产品在室温或2-8℃下可稳定保存24个月，储存、运输和操作都更为便捷。37℃保存2周，酶活下降不超过10%，而使用的重组猪胰蛋白酶的消化液在37℃保存条件下非常不稳定。
注意事项：
虽然本产品在室温非常稳定，但仍建议4℃保存。如果室温保存，需要避免阳光直射，并尽量避免光照，同时需要确保室温在适当范围内，避免出现30℃以上室温长时间放置。
本产品从4℃取出后可直接用于细胞的消化。对于较难消化的细胞或需要快速消化的情况，在加入本产品后，可将培养器皿置于37℃温育1-3分钟。
在使用本产品的过程中要特别注意避免消化液被微生物污染。
本产品长期不使用也可-20℃保存。虽然经测试，反复冻融5次对酶活几乎没影响，但还是需要尽量避免反复冻融。
本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。
为了您的安全和健康，请穿实验服并戴一次性手套操作。
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Aladdin 's EnzymoPure™ Express Enzyme is an animal origin-free (AOF) and ready-to-use cell dissociation solution containing a recombinant trypsin that is stable at room temperature. This product digests cells mildly, rapidly, and effectively, with almost no damage to cells, and can be a perfect substitute for ordinary trypsin cell digestion solution.
This product is sterilized by filtration and contains approximately 1mM EDTA, with a pH value of 7.2-7.8. Not only can it be used directly to digest cultured cells or some tissues, but it is more suitable for digestion of cells or tissues that strictly require the absence of animal origin.
Trypsin is a serine protease that specifically cleaves peptide bonds at the carboxyl side of lysin or arginine residues. Aladdin's EnzymoPure™ Express Enzyme employs a high-purity recombinant trypsin-like enzyme, also known as Trypsin replacement, with a fungal-derived protein sequence and the same enzymatic properties as porcine-derived trypsin, but with higher stability, better purity, milder digestion and lower cytotoxicity, which can be used as a substitute for porcine pancreatic-derived trypsin for cell and tissue dissociation and other applications.
The trypsin replacement in this product is not of animal origin. This product is free of animal-derived viral contamination as well as other animal-derived impurities. Both the amounts of host protein and DNA residues in this product meet the pharmacopoeia standards.
This product has good digestion effect. Under the same conditions, the enzymatic activity of this product is approximately 2 times higher than that of ordinary cell digestion solution containing 0.05% trypsin, and it is more effective for most cell types. Usually most of the adherent cells can be digested within 1-3 minutes at room temperature. Please refer to Figure 1 for the cell digestion effect of this product. Cells can usually be blown off when there is a visible slight morphological change and before the cells are completely rounded.
Figure 1. Digestion of A498, BEAS-2B, and L929 cells with Aladdin's EnzymoPure™ Express Enzyme. In this experiment, this product placed at room temperature for about 10 minutes was directly added to cells after aspirating cell culture medium. Within 1-2 minutes, treated cells demonstrated contracted edges and gradually dissociated into single cells. Pictures were taken after different time courses as indicated. The digestion effect will be better if cells are washed with PBS or HBSS before digestion. This figure is for reference only, which may vary due to different experimental conditions.
This product utilizes a high-purity recombinant trypsin replacement that has high enzyme specific activity, reducing the risk of cellular damages caused by other enzymes. It can minimize the cellular damages by over-digestion (Figure 2), eliminating the need for enzyme inhibitors or serum to terminate the reaction.
Figure 2. Growth state of cells digested with Aladdin's EnzymoPure™ Express Enzyme. A498 and BEAS-2B cells were digested with EnzymoPure™ Express Enzyme, respectively. Cells were over-digested until they were all floating and no trypsin inhibitor was used to terminate the reaction. Over-digested cells was collected by centrifugation and cultured for 18 h after re-plating. No significant differences between the EnzymoPure™ over-digested cells and the normal digested cells were observed. This figure is for reference only, which may vary due to different experimental conditions.
Precautions：
Although this product is very stable at room temperature, we still recommend storing it at 4℃. If stored at room temperature, it should be kept in the dark. Room temperature should be within an appropriate range and prolonged storage at room temperature over 30℃ should be avoided.
This product can be used directly for cell digestion after removal from 4℃. For cells that are more difficult to digest or where rapid digestion is required, digestion can be performed at 37℃ for 1-3 minutes.
Special care should be taken to avoid microbial contamination of this product.
This product can be stored at -20℃ for long-term storage. Avoid repeated freeze-thaw.
This product is for R&D only. Not for drug, household, or other uses.
For your safety and health, please wear a lab coat and disposable gloves during the operation.
规格
产品名称
EnzymoPure™ Express Enzyme
分子类型
酶
储存与运输
储存条件
2-8°C储存
运输条件
低温运输
稳定性与储存
室温或4℃保存，两年有效。4℃保存，效果更好。
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产品仅提供给经过验证的企业、机构和合格的专业人员用于研发。不适用于人类、动物、诊断或治疗。
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