计算溶液所需的质量、体积或浓度。
外泌体提取纯化试剂盒 (体液)
有货
为什么选择此级别
BioReagent,无菌 级 ，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。常规运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
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
外泌体是由细胞分泌的包含 RNA 和蛋白质的小囊泡 （30-150 nm），在血液、唾液、尿液及乳汁等体液中大量存在。外泌体被认为具有细胞间信使的功能，在特定细胞之间传递它们的效应物或信号分子；然而其构造、效应物组成 以及所参与的生物学通路目前尚不明晰。外泌体的生物学功能研究中需要分离完整的外泌体颗粒，而传统超速离心方法步骤繁琐、硬件要求高、操作难度大。该外泌体提取纯化试剂盒，组分经过优化处理，适用于脑脊液、羊水、乳汁、唾液、肺泡灌洗液以及鼻腔盥洗液等体液中的外泌体提取，并搭配纯化过滤装置，可快速高效地获得高纯度外泌体颗粒，可用于电镜分析、NTA 粒径分析、核酸分析、蛋白分析、细胞学实验和动物实验等。
产品组成
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自备材料：
高速离心机，涡旋振荡器，50 mL 离心管，1.5 mL 离心管，1×PBS 缓冲液（无菌）。
操作规程：
一、 样品预处理
1、 取样：如果是冻存样品，从冰箱取出后于 25℃水浴中进行解冻，将完全融化后的样品置于冰上，如果是新鲜样品，收集样品后置于冰上；
2、 样品初始用量：单次提取时的样品量建议肺泡灌洗液、腹水等不低于 1 mL，建议 1 次提取 2-3 mL 体积；
3、 离心去细胞碎片：将样品转移至离心管中，于 4℃以3,000 ×g（~6,200 rpm*）离心 10 min，去除样品中的细胞碎片（注：若沉淀较多，可 3,000 ×g，10 min离心多次至无明显沉淀，每次取离心上清液）；*为约7 cm有效离心半径的小离心机换算（≤2 mL离心管），下同。
4、 离心去杂质碎片：将离心上清液转移至新的离心管中，于 4℃以 12,000 ×g（~12,400 rpm）离心 10 min，去除样品中的杂质碎片；
5、 上清液转移：去除杂质碎片的上清液转移到新离心管中。
二、 提取外泌体
1、 上清液预处理：在去除杂质的离心上清液中加入外泌体浓缩液（ECS 试剂），具体的加入剂量如下（其他剂量请根据表中的用量等比例换算）：
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2、 溶液混合：加入 ECS 试剂后将离心管盖紧，通过涡旋振荡器混匀 1 min，再放置于 4℃静置８ h 以上（注：增加静置时间可提高外泌体得率，但不可超过 24 h）；
3、 沉淀外泌体：取出装有混合液的离心管于 4℃以 12,000 ×g（~12,400 rpm）离心 30 min，弃上清，沉淀中富含外泌体颗粒（尽可能吸尽上清液）；
4、 再次离心：将含有沉淀的离心管再次于 4℃以 12,000 ×g（~12,400 rpm）离心 2 min，弃上清（注：尽可能吸尽上清液）；
5、 外泌体重悬：取合适量的 1×PBS 均匀吹打离心沉淀物，待其溶解后，将重悬液转移至新的 1.5 mL 离心管中（建议每 1 mL 体液用 100 μL 左右 1×PBS 重悬）；
6、 收获外泌体颗粒：将含有重悬液的1.5 mL 离心管于4℃以 12,000 ×g（~12,400 rpm）离心 2 min，保留上清液，其中富含外泌体颗粒（注：若沉淀较多，可 12,000×g，2 min 离心多次至无明显沉淀，每次取上清液）。
三、 纯化外泌体
1、 纯化外泌体：将收获的同一体液来源的外泌体颗粒粗品合并转入外泌体纯化滤器（EPF柱）上室中，于 4℃以 3,000 ×g（~6,200 rpm）离心 10 min，离心后收集 EPF 柱管底的液体，此液体即为纯化后的外泌体颗粒（注：EPF 柱不可重复使用）；
2、 外泌体的保存：纯化后的外泌体以合适体积进行分装冻存于-80℃低温冰箱中，以备后续实验使用。
注意：
本产品仅用于生命科学研究，不得用于医学诊断及其他用途！
Exosomes are small vesicles (30–150 nm) secreted by cells, containing RNA and proteins, and are abundantly present in body fluids such as blood, saliva, urine and milk. Exosomes act as intercellular messengers to deliver effector molecules or signaling molecules between specific cells. Nevertheless, their structure, composition of effector substances and the biological pathways they participate in remain poorly characterized. Intact exosome particles are indispensable for research on the biological functions of exosomes. Conventional ultracentrifugation is plagued by complicated workflows, high requirements for specialized equipment and cumbersome operation. This exosome isolation and purification kit adopts optimized formulations, which is applicable to exosome extraction from body fluids including cerebrospinal fluid, amniotic fluid, milk, saliva, bronchoalveolar lavage fluid and nasal lavage fluid. Equipped with a purification filter device, the kit enables rapid and efficient acquisition of high-purity exosome pellets. The purified products can be applied to transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), nucleic acid analysis, protein analysis, cell-based assays and animal experiments.
Product Components
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Required Materials
High-speed centrifuge, vortex mixer, 50 mL centrifuge tubes, 1.5 mL centrifuge tubes, sterile 1×PBS buffer.
Operating Protocol
I. Sample Pretreatment
1.Sample handling and thawing: For frozen samples, take them out from the refrigerator and thaw in a 25 °C water bath, then place fully thawed samples on ice. Fresh samples shall be placed on ice immediately after collection.
2.Recommended initial sample volume: For single isolation, the minimum recommended volume for bronchoalveolar lavage fluid, ascites and other body fluids is 1 mL; 2–3 mL is preferred per isolation run.
3.Centrifugation to remove cell debris: Transfer samples into centrifuge tubes and centrifuge at 3,000 ×g (~6,200 rpm*) for 10 min at 4 °C to remove cell debris. Note: If abundant pellets are observed, repeat centrifugation at 3,000 ×g for 10 min until no obvious precipitate exists, and retain the supernatant after each round of centrifugation.
*Calculated for small centrifuges with an effective radius of approximately 7 cm (for tubes ≤2 mL), the same applies hereinafter.
4.Centrifugation to remove impurity fragments: Transfer the supernatant to a new centrifuge tube and centrifuge at 12,000 ×g (~12,400 rpm) for 10 min at 4 °C to eliminate residual impurities and debris.
5.Supernatant transfer: Transfer the clarified impurity-free supernatant into a new centrifuge tube.
II. Exosome Isolation
1.Supernatant pretreatment: Add Exosome Concentration Solution (ECS reagent) to the clarified supernatant. Refer to the table below for detailed dosing standards; volumes for other sample input amounts can be calculated proportionally based on the listed ratios.
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2. Solution Mixing: After adding ECS reagent, tightly cap the centrifuge tube and vortex thoroughly for 1 minute on a vortex mixer. Incubate the mixture at 4 °C for no less than 8 hours. Note: Extended incubation time can improve exosome yield, but the incubation duration must not exceed 24 hours.
3. Exosome Precipitation: Take out the tube containing the mixture and centrifuge at 12,000 ×g (~12,400 rpm) for 30 minutes at 4 °C. Discard the supernatant completely to the greatest extent possible; the pellet is enriched with exosome particles.
4. Secondary Short Spin: Centrifuge the tube with the pellet again at 12,000 ×g (~12,400 rpm) for 2 minutes at 4 °C, then thoroughly remove all residual supernatant.
5. Exosome Resuspension: Gently pipette the pellet up and down with an appropriate volume of 1×PBS buffer until fully dissolved, then transfer the resuspension to a new 1.5 mL centrifuge tube. Recommended resuspension volume: approximately 100 μL of 1×PBS for every 1 mL starting body fluid sample.
6. Harvest Crude Exosome Pellets: Centrifuge the 1.5 mL tube with resuspended sample at 12,000 ×g (~12,400 rpm) for 2 minutes at 4 °C, retain the supernatant which contains enriched crude exosomes. Note: If visible precipitate remains, repeat centrifugation at 12,000 ×g for 2 minutes until no obvious pellet is observed, and collect the supernatant after each centrifugation.
III. Exosome Purification
1. Column Purification: Combine all crude exosome suspensions derived from the same type of body fluid and load them into the upper chamber of the Exosome Purification Filter (EPF column). Centrifuge at 3,000 ×g (~6,200 rpm) for 10 minutes at 4 °C. Collect the eluate at the bottom of the EPF column, which is the final purified exosome product. Note: EPF columns are disposable and cannot be reused.
2. Exosome Storage: Aliquot the purified exosomes into appropriate volumes and store long-term in a -80 °C freezer for subsequent downstream experiments.
Note:
This product is for life science research use only. It shall not be applied for medical diagnosis or any clinical purposes.
规格
别名
通用体液外泌体提取纯化试剂盒
英文别名
Body Fluid-Derived Exosome Extraction & Purification Kit
规格或纯度
BioReagent, 无菌
英文名称
Exosome Isolation and Purification Kit (Biofluids)
应用
Exosome Isolation and Purification
样品类型
体液
储存与运输
储存条件
室温
运输条件
常规运输
稳定性与储存
Store at RT long term (24 months).
组分与储存
| 组分货号 | 组分名称 | 物理外观 | 3T | 储存 |
| E1526241A | 外泌体浓缩液 | 液体 | 1.8 mL | RT |
| E1526241B | 外泌体纯化滤器 | — | 3 EA | RT |
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
通过匹配包装上的批号来查找并下载产品的 COA，每批产品都进行了严格的验证，您可放心使用！
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0939125 |
分析证书 | E1526241 |
技术文档和文章
溶液计算器
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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