计算溶液所需的质量、体积或浓度。
外泌体提取纯化试剂盒 (多合一)
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，适用于细胞培养上清液、血清血浆中的外泌体提取，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细胞培养上清液不低于 20 mL，血清/血浆不低于 0.5 mL；
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*/~5,200 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。**为约10 cm有效离心半径的大离心机换算（≥15 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 10,000 ×g（~11,300 rpm*/~9,500 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），部分较为粘稠的样品（血清、血浆等），需先加入预冷的1×PBS 进行稀释，然后再加入 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 10,000 ×g（~11,300 rpm*/~9,500 rpm**）离心 60 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 10,000 ×g（~11,300 rpm*/~9,500 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 20 mL 细胞培养上清液或 0.25 mL 血清/血浆用200 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：将收获的外泌体颗粒粗品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 that contain RNA and proteins, and they are abundant in body fluids such as blood, saliva, urine and milk. Exosomes are regarded as intercellular messengers that deliver effector molecules or signaling molecules between specific cells. Nevertheless, their structure, composition of effector substances and involved biological pathways remain poorly understood.
Intact exosome particles are required for research on the biological functions of exosomes. Conventional ultracentrifugation involves complicated procedures, demands specialized equipment and is difficult to operate. This exosome isolation and purification kit features optimized components. Compatible with exosome extraction from cell culture supernatant, serum and plasma, it is equipped with a purification filter device to rapidly and efficiently obtain high-purity exosome particles. The isolated exosomes 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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Materials to Be Prepared
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 collection and thawing: For frozen samples, take them out of the refrigerator and thaw in a 25 °C water bath, then place fully thawed samples on ice. Fresh samples should be placed on ice immediately after collection.
2.Recommended initial sample volume: For single isolation, the minimum recommended volume is 20 mL for cell culture supernatant and 0.5 mL for serum/plasma.
3.Centrifugation to remove cell debris: Transfer samples into centrifuge tubes and centrifuge at 3,000 ×g (~6,200 rpm* / ~5,200 rpm**) for 10 min at 4 °C to eliminate cell debris. Note: If a large pellet is observed, repeat centrifugation at 3,000 ×g for 10 min until no obvious precipitate remains, and retain the supernatant after each centrifugation.
* Calculated for small centrifuges with an effective radius of approximately 7 cm (for tubes ≤2 mL), same hereinafter.
** Calculated for large centrifuges with an effective radius of approximately 10 cm (for tubes ≥15 mL), same hereinafter.
4.Centrifugation to remove impurity fragments: Transfer the supernatant to a new centrifuge tube and perform a second centrifugation at 10,000 ×g (~11,300 rpm* / ~9,500 rpm**) for 10 min at 4 °C to remove residual impurities and fragments.
5.Supernatant transfer: Transfer the clarified supernatant free of impurities into a new centrifuge tube.
II. Exosome Isolation
1.Supernatant pre-treatment: Add Exosome Concentration Solution (ECS reagent) to the clarified supernatant. For viscous samples such as serum and plasma, dilute the sample with pre-cooled sterile 1×PBS prior to adding ECS reagent. Refer to the table below for detailed dosing ratios; volumes for other input amounts can be calculated proportionally based on the listed dosages.
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2. Solution Mixing: After adding ECS reagent, tightly cap the centrifuge tube, vortex for 1 minute on a vortex mixer, then incubate the mixture at 4 °C for no less than 8 hours. Note: Longer incubation time can improve exosome yield, but the incubation period shall not exceed 24 hours.
3. Exosome Precipitation: Take out the tube containing the mixture and centrifuge at 10,000 ×g (~11,300 rpm* / ~9,500 rpm**) for 60 minutes at 4 °C. Discard the supernatant completely as far as possible; the pellet is enriched with exosome particles.
4. Secondary Short Spin: Centrifuge the tube with the pellet again at 10,000 ×g (~11,300 rpm* / ~9,500 rpm**) for 2 minutes at 4 °C, then thoroughly remove the 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 200 μL of 1×PBS for every 20 mL cell culture supernatant or 0.25 mL serum/plasma starting material.
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, collect supernatant each time.
III. Exosome Purification
1. Column Purification: Load the crude exosome suspension 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 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 at -80 °C freezer for subsequent downstream experiments.
Notice: This product is for research use only in life science fields. It shall not be used for medical diagnosis or any clinical applications.
规格
别名
一体式外泌体分离纯化试剂盒
英文别名
All-in-One Exosome Extraction & Purification Kit
规格或纯度
BioReagent, 无菌
英文名称
Exosome Isolation and Purification Kit (All in one)
应用
Exosome Isolation and Purification
样品类型
细胞, 血浆, 血清
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 | 2T | 储存 |
| E1526251A | 外泌体浓缩液 | 液体 | 12 mL | RT |
| E1526251B | 外泌体纯化滤器 | — | 2 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 |
|---|---|---|
 ZJ26F0939126 |
分析证书 | E1526251 |
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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