计算溶液所需的质量、体积或浓度。
为什么选择此级别
，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
室温。 。请查阅批次 COA 获取详细规格。
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文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
本产品专为人、小鼠、大鼠等动物骨骼肌组织单细胞悬液制备设计，通过多元复合酶的温和酶解作用将骨骼肌组织高效解离成为单个细胞悬液，在高得率的同时能够获得高活性的单细胞，后续适用于单细胞测序、流式细胞分析、流式细胞分选、原代细胞培养、类器官 3D 培养等实验。
组分表：
优势特点
1、一步即用型
本产品只需把配套的酶干粉溶解在溶解液里，一步操作即可进行后续实验。无需像进口竞品一样，把三四种酶分别溶解在不同溶液里，再分别进行体积计算，后量取不同体积的酶混合在一起并混匀才能使用，本产品无需任何称量、计算，避免多步骤操作和复杂的计算，减少出错概率。
2、多元复合酶
本产品是自主研发生产的多元复合酶，根据不同组织的细胞间质种类配备 8 种以上的复合酶，可以更好的消化解离细胞间的各种组织基质，比单纯使用进口胶原酶 IV、DNA 酶或进口竞品的三四种混合酶的解离效率更高，解离时间短、细胞得率高、细胞活性强、抗原表位完整，自研自产成本更低，更有竞争力。
3、解离时间短
由于本产品使用的是 8 种以上自研自产的特异性组织解离酶，大多数器官组织在 10-15 分钟即可解离完全，解离时间超短，而单纯使用进口胶原酶 IV、DNA 酶或进口竞品的三四种混合酶需要 1 小时左右，甚至需要 2-3 小时才能解离完全。
4、细胞得率高
建立在多元复合酶的基础上，该产品可以更好的消化细胞间的各种组织基质，组织消化更彻底，组织单细胞完全释放出来，组织团块残留少，因此细胞得率更高。
5、细胞活性强
针对不同的组织该产品配套了不同的特异性解离复合酶，该复合酶是温和的解离方式，不像胰酶一样对细胞有较大损伤，并且该复合酶酶解消化时间更短，大幅减少了细胞的离体时间，防止了解离酶的过度消化，因此能得到更高活率的单细胞、抗原表位更完整的细胞。
6、抗原表位完整
由于该产品酶解效率更高，酶解时间和细胞离体时间更短，组织细胞损伤更低，细胞结构完整性更好，细胞活性更强，因此抗原表位更完整，更有利于抗体结合染色，解离的单细胞更适合进行流式分析分选实验。
使用方法
1、溶解酶干粉
首次启用试剂盒时执行该操作步骤。将 A 瓶中的酶干粉用力甩至瓶底；然后用移液枪取 B 瓶中的溶解液反复吹打 A 瓶中的酶干粉并转移至 B 瓶中，必要时可将枪头口用剪刀剪大，确保 A 瓶中酶干粉全部转移至 B 瓶中；随后盖紧 B 瓶瓶盖上下颠倒 B 瓶直至酶干粉完全溶解，必要时可放置摇床上摇晃至完全溶解，完全溶解的酶溶液呈现清澈微棕色。
2、过滤分装解离酶
在 A 瓶中酶干粉完全溶解在 B 瓶溶解液后，根据后续实验如需要无菌解离组织，则需用注射器和 0.22μm 无菌过滤器过滤除菌后分装。建议每 5mL 分装一瓶，分装至 A 瓶中，分装后应立即冻存在 - 20℃或 - 80℃冰箱中，可保存 6 个月。单次未使用完可冻存后下次继续使用，但反复冻融不要超过三次，每冻融一次酶活会降低一部分，酶活降低时可增加酶的使用量以增强酶解效果。
3、酶解实验操作
(1) 实验准备：预制碎冰，冰上解冻单细胞解离酶和酶解终止液，单细胞解离仪器开机预热，PBS 预冷，眼科剪、镊子等无菌手术器械，40μm 无菌细胞筛，离心管、离心机等预冷。
(2) 组织收集：无菌条件下收集组织，立即置于冷的 PBS 中，尽可能 4h 内处理，如需放置更长时间，请使用高活性组织保存液。
(3) 清洗组织：将组织用冷 PBS 洗 1-2 次以去除血液、坏死组织等杂质，称重记录。
(4) 剪碎组织：将清洗过的组织转移至解离管中，用锋利眼科剪碎成 1-2mm³ 小块，剪至泥糊状，剪得越碎酶解就越充分，单细胞酶解时间就会越短；如有配备组织单细胞解离仪，觉得剪碎较麻烦也可不用严格剪碎，同样能获得较好的解离效果。
(5) 酶解组织：组织剪碎后，取适量单细胞解离酶加入解离管中，上下颠倒解离管使解离酶与组织充分混匀。
① 如配备有组织单细胞解离仪，可按照仪器操作规范进行单细胞解离。本产品适用市面上所有进口或国产品牌的组织单细胞解离仪，但需注意本产品解离效率比自备酶或其他竞品的酶都高，解离时间可减少至少一半，需每隔 5-10min 确认一下解离情况，不同组织解离完全的时间从 5min 到 50min 不等，在保证解离效果情况下尽可能减少酶解时间，酶解完全即可终止酶解；
② 如未配备组织单细胞解离仪，也可进行手动解离。在剪碎组织步骤中需严格按照要求剪碎组织；根据组织和所加酶体积选择在 1.5mL EP 管或 5mL EP 管中，将解离酶与组织充分混匀，用剪刀将 1mL 移液枪枪头口稍微剪大后，用移液枪用力吹打混合物，如吹打堵塞枪头说明组织剪碎不够彻底，可继续剪碎组织或将枪头再剪大一些，保证能用移液枪反复吹打混合物，反复吹打十余次后立即放入 37℃培养箱、水浴或金属浴中孵育，有摇床效果更佳，每孵育 5 分钟需进行一次反复吹打，直至组织团块完全解离成单细胞悬液。
(6) 单细胞过滤：待组织完全解离后，立即瞬离解离管 3s，并将解离管插在冰上，用移液枪吸取上清通过 40μm 的滤网进行过滤，过滤全程在冰上操作。
(7) 终止酶解：取等量的酶解终止液或完全培养基进行终止酶解。
(8) 收集细胞：离心 5 分钟后弃上清。
(9) 红细胞裂解： 取适量红细胞裂解液重悬细胞沉淀，冰上孵育 10 分钟，间歇吹打轻摇，4℃、300g 离心后弃上清。
(10) 清洗：4℃离心后弃上清。
(11) 去除碎片： 如碎片较多，可用高效碎片去除剂去除碎片。
(12) 细胞活性检测：用台盼蓝或 AOPI 染色检测，活细胞率应 > 85%。
(13) 细胞冻存：如单细胞解离后暂时无法进行后续实验，可使用高效高活细胞冻存液将单细胞重悬后直接冻存于 - 80℃，可冻存数年。
注意事项
本产品仅供科研使用；如下游实验需无菌培养，全程需在超净台内进行无菌操作，避免污染；酶解需在 37℃，其他操作均在冰上操作，严格控制温度；根据组织酶解量调整酶解时间，勤观察及时确认裂解阶段，在保证裂解效果情况下尽可能减少酶解时间，以免时间过长造成对细胞的损伤；若用于单细胞测序等要求较高的单细胞制备，全程需加快速度，单细胞解离后需立即进入下游建库步骤；该酶溶解后 - 20℃或 - 80℃保存，可保存 6 个月，单次未使用完可冻存后下次继续使用，但反复冻融不要超过三次，每冻融一次酶活会降低一部分，酶活降低时可增加酶的使用量以增强酶解效果。
This product is specifically designed for preparing single-cell suspensions from skeletal muscle tissues of humans, mice, rats, and other animals. Through the gentle enzymatic hydrolysis of a multi-component composite enzyme, cardiac tissues are efficiently dissociated into single-cell suspensions, achieving high yield while maintaining high cell viability. The resulting single cells are suitable for downstream experiments including single-cell sequencing, flow cytometry analysis, flow cytometric sorting, primary cell culture, and organoid 3D culture.
Component List
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Product Advantages
1. Ready-to-Use (One-Step Operation)
Simply reconstitute the provided enzyme powder with the supplied buffer for immediate use in downstream experiments. Unlike imported competitors that require separate reconstitution of 3-4 enzymes in different solutions, volume calculation, and mixing, this product eliminates the need for weighing, complex calculations, and multi-step operations—significantly reducing the risk of errors.
2. Multi-Component Composite Enzyme
Independently developed and manufactured, this product contains over 8 types of composite enzymes tailored to the extracellular matrix components of different tissues. It achieves more efficient digestion of intercellular matrices compared to single-use collagenase IV, DNase, or 3-4 component mixed enzymes from imported brands. Key benefits include shorter digestion time, higher cell yield, superior cell viability, intact antigen epitopes, and more competitive pricing due to independent R&D and production.
3. Short Digestion Time
Leveraging over 8 types of self-developed tissue-specific dissociation enzymes, most organ tissues are fully dissociated within 10-15 minutes. In contrast, single collagenase IV, DNase, or imported mixed enzymes typically require ~1 hour, and sometimes 2-3 hours, for complete dissociation.
4. High Cell Yield
The multi-component composite enzyme ensures thorough digestion of intercellular matrices, maximizing the release of single cells with minimal tissue clump residues—resulting in higher overall cell yield.
5. Superior Cell Viability
Tissue-specific composite enzymes enable gentle dissociation, causing less cell damage than trypsin. Combined with shorter digestion time, the product minimizes in vitro cell exposure and avoids over-digestion, preserving high cell viability and intact antigen epitopes.
6. Intact Antigen Epitopes
Efficient digestion reduces both processing time and in vitro cell exposure, minimizing tissue/cell damage while maintaining structural integrity and high viability. Intact antigen epitopes facilitate antibody binding and staining, making the dissociated single cells ideal for flow cytometry analysis and sorting.
Usage Instructions
1. Reconstitute Enzyme Powder
Perform this step when first using the kit:
Centrifuge the vial of Enzyme Powder A briefly to collect all powder at the bottom.
Use a pipette to transfer Reconstitution Buffer B into the vial of Enzyme Powder A, repeatedly pipetting to resuspend the powder. For easier transfer, trim the tip of the pipette if necessary to ensure complete transfer of the powder into Buffer B.
Cap the vial tightly and invert to mix until the powder is fully dissolved (use a shaker if needed). The fully reconstituted enzyme solution will appear clear and slightly brown.
2. Filter and Aliquot Dissociation Enzyme
After complete reconstitution:
For sterile tissue dissociation (e.g., cell culture), filter the enzyme solution through a 0.22μm sterile filter using a syringe, then aliquot.
Recommended aliquot volume: 5mL per vial (reuse vial A if desired).
Store aliquots at -20℃ or -80℃ immediately; stable for 6 months. Unused aliquots can be refrozen for future use, but avoid more than 3 freeze-thaw cycles (each cycle reduces enzyme activity—adjust volume accordingly to compensate for reduced activity).
3. Enzymatic Digestion Procedure
(1) Pre-Experiment Preparation
Prepare crushed ice; thaw the single-cell dissociation enzyme and digestion stop solution on ice.
Preheat the single-cell dissociation instrument (if used); pre-cool PBS, sterile surgical tools (ophthalmic scissors, forceps), 40μm sterile cell strainers, centrifuge tubes, and centrifuge.
(2) Tissue Collection
Collect tissues under sterile conditions and immediately place in cold PBS. Process within 4 hours if possible; for longer storage, use High-Activity Tissue Preservation Solution.
(3) Tissue Washing
Wash tissues 1-2 times with cold PBS to remove blood, necrotic tissue, and other impurities. Weigh and record the tissue mass.
(4) Tissue Mincing
Transfer washed tissues to a dissociation tube and mince into 1-2mm³ pieces (mush-like consistency) using sharp ophthalmic scissors. Finer mincing ensures more efficient digestion and shorter processing time. If using a single-cell dissociation instrument, mincing can be less rigorous while maintaining good dissociation results.
(5) Tissue Digestion
Add an appropriate volume of the reconstituted dissociation enzyme to the tube and invert to mix thoroughly with the minced tissue.
① If a tissue single-cell dissociation instrument is available, perform single-cell dissociation according to the instrument's operating specifications. This product is compatible with all commercial tissue single-cell dissociation instruments (both imported and domestic brands). However, note that its dissociation efficiency is higher than that of user-supplied enzymes or enzymes from other competitors, enabling at least a 50% reduction in dissociation time. Check the dissociation progress every 5-10 minutes—complete dissociation time varies by tissue (ranging from 5 to 50 minutes). Minimize enzymatic digestion time as much as possible while ensuring dissociation efficiency, and terminate digestion immediately once complete.
② If no tissue single-cell dissociation instrument is available, manual dissociation is also feasible:
Ensure tissues are minced strictly as required in the tissue mincing step.
Select a 1.5mL or 5mL EP tube based on the tissue quantity and enzyme volume added. Thoroughly mix the dissociation enzyme with the minced tissue.
Slightly trim the tip of a 1mL pipette with scissors, then vigorously pipette the tissue-enzyme mixture. If pipetting is blocked, the tissue is insufficiently minced—re-mince the tissue or trim the pipette tip further to ensure smooth repeated pipetting.
After 10+ rounds of vigorous pipetting, immediately incubate the tube at 37℃ in an incubator, water bath, or dry bath (a shaking incubator is preferred).
Vigorously pipette the mixture every 5 minutes until all tissue clumps are fully dissociated into a single-cell suspension.
(6) Single-Cell Filtration
Once dissociation is complete, centrifuge the tube briefly (3 seconds) and place on ice.
Pipette the supernatant through a 40μm cell strainer into a new tube (perform all steps on ice).
(7) Stop Digestion
Add an equal volume of Digestion Stop Solution or complete medium to terminate digestion.
(8) Cell Collection
Centrifuge for 5 minutes and discard the supernatant.
(9) Red Blood Cell (RBC) Lysis
Take an appropriate volume of red blood cell lysis buffer to resuspend the cell pellet.
Incubate on ice for 10 minutes, gently pipetting intermittently.
Centrifuge at 300g for 5 minutes at 4℃ and discard the supernatant.
(10) Cell Washing
Centrifuge at 4℃ and discard the supernatant.
(11) Debris Removal
If there is a high amount of debris, a high-efficiency debris remover can be used to eliminate it.
(12) Cell Viability Assessment
Stain with trypan blue or AOPI; viable cell rate should exceed 85%.
(13) Cell Cryopreservation
If downstream experiments are delayed, resuspend single cells in High-Efficiency High-Viability Cell Cryopreservation Solution and store directly at -80℃ (stable for years).
Notes
For research use only.
For sterile culture applications, perform all operations in a biosafety cabinet to avoid contamination.
Digestion must be conducted at 37℃; all other steps should be performed on ice to strictly control temperature.
Adjust digestion time based on tissue quantity. Monitor dissociation progress closely and stop digestion as soon as complete to prevent cell damage.
For high-demand applications (e.g., single-cell sequencing), accelerate the entire process and proceed immediately to library preparation after dissociation.
Store reconstituted enzyme at -20℃ or -80℃ (stable for 6 months). Unused aliquots can be refrozen, but avoid more than 3 freeze-thaw cycles (each cycle reduces activity—increase enzyme volume if needed to maintain digestion efficiency).
规格
产品名称
骨骼肌组织单细胞解离酶
分子类型
酶
产品线
生物产品
储存与运输
储存条件
室温
稳定性与储存
长期储存2-8℃（12个月）。 溶解后 -20℃ 储存。
技术文档
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0130968 |
分析证书 | S1507385 |
技术文档和文章
溶液计算器
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