计算溶液所需的质量、体积或浓度。
细胞上清/尿液外泌体纯化试剂盒 (SEC)
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
2-8°C储存。低温运输 。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
外泌体纯化试剂盒（SEC）是基于尺寸排阻色谱法（Size Exclusion Chromatography, SEC）原理的外泌体纯化试剂盒。色谱柱采用稳定、多孔的凝胶颗粒填充，将含有外泌体的溶液加入到SEC色谱柱中时，较小的颗粒将被捕获在孔中，而较大的分子则不会进入孔中并被率先洗脱。因此，不同阶段的洗脱部分将包含不同大小的分子，首先是较大的外泌体颗粒，其次是较小的蛋白质和脂质颗粒。本试剂盒更适合用于下游为外泌体功能验证、表征、内容物的检测（核酸，蛋白，脂质和代谢物检测或组学分析）等的研究。
组分与储存：
| 组分货号 | 组分名称 | 物理外观 | 1ml | 储存 |
| C778164A | SEC纯化柱 (1mL) | 液体 | 1 mL | 2-8℃ |
| C778164B | 保存液 (5×) | 液体 | 30 mL | 2-8℃ |
| C778164C | 清洗液 (5×) | 液体 | 20 mL | 2-8℃ |
产品特点：
纯度高：杂质蛋白去除率高达99.44%，从源头保障后续研究的准确性。
方便快捷：操作简单，整个操作仅需30min。
结果稳定：重复性高，实验结果高度吻合。
产品原理：
图1. 外泌体纯化试剂盒（SEC）作用原理
产品数据：
1：NTA检测结果
图2. NTA结果显示，外泌体主要存在于3-7组分
2：TEM检测结果
图3. TEM结果显示，经过SEC柱2.0纯化收集的外泌体，形态完整且清晰
3：BCA蛋白检测结果
图4. BCA结果显示，杂蛋白去除率高达99.5%以上
操作流程：
样本预处理：
1.细胞上清
①收集细胞上清10-50mL（收集时细胞培养密度不低于1X10⁶cfu/mL）；
②细胞上清离心（3000g、10min），保留上清；
③上清样本使用0.22μm滤膜过滤；
④ 过滤后用100kD超滤管进行浓缩，浓缩体积比为1:20，收集浓缩液备用。
2.尿液
①采集中段晨尿约10-50mL，离心（3000g、10min）， 保留上清；
② 上清液使用0.22μm 滤膜过滤；
③ 过滤后用100kD超滤管浓缩，浓缩体积比为1:20，收集浓缩液备用。
外泌体分离：
1. 试剂准备
保存液(5×)和清洗液(5×)均需要用去离子水稀释到1×，所用试剂需用0.22μm滤膜过滤后再使用；
自备1×PBS，使用0.22μm滤膜过滤后再使用或者采购已过膜的PBS（使用前需平衡到室温）。
2. 柱平衡
① 从冰箱取出SEC纯化柱，将其垂直固定（需自备固定装置）；
② 置于室温下放置30分钟；
③ SEC纯化柱下方放置一个空烧杯；
④ 打开顶部柱塞和底盖，将管中保存液排空至底部出口无液体流出，向柱中加入15mL 1×PBS，可分次加，让液体全部流出，再上样。
3. 样品上样
① 取8个EP管，依次标记为 1，2，3，4，5，6，7，8；
② 将1mL样品从SEC纯化柱顶部沿壁加入，样品完全进入填料并且底部出口无液体流出后，加入0.5 mL 1×PBS，使用EP管1收集流出液，收集体积为0.5mL；
③ 待上一个0.5 mL 1×PBS全部进入排阻柱内后，再添加0.5 mL 的 1×PBS，使用EP管2收集流出液，收集体积为0.5 mL；
注意：使用时应尽量保证柱内有液体，以确保填料能够被充分润湿。
④ 重复③操作6次，总共收集8管洗脱液。
注意：1. 样品最小体积不应小于1mL；样品不足1mL时，用1×PBS补到1mL，混匀后再上样。
4. 外泌体收集
① 若想获得高回收率外泌体：保留 3，4，5，6，7 的馏分，共计2.5mL；若不区分馏分，在第2个0.5 mL 1×PBS全部进入排阻柱内 后，直接加入2.5 ml 1×PBS，洗脱至一个收集管中。
② 若想获得高纯度外泌体：将第 4，5，6 的馏分合并，共计1.5mL。
注意： 如需浓缩外泌体洗脱液，可使用100 kDa 超滤管，4000g 离心浓缩到目的体积即可。
5. SEC纯化柱的清洗与储存
收集结束后，加10mL清洗液，接着用1×PBS 冲洗15mL，再用1×保存液冲洗10mL。将底盖关闭，用1×保存液填满SEC纯化柱，用柱塞封闭顶部端口并检查无液体流出后，4 ℃保存。
注意：1. 该SEC纯化柱可重复使用10次。
2. 使用结束后需要用封口膜将柱上端封死，避免保存液挥发。
免责说明：
本品仅限于科学研究使用，不得用于临床诊断或治疗，不得用于食品或药品。
Exosome Purification Kit (SEC) is developed based on the principle of Size Exclusion Chromatography (SEC). The chromatographic column is packed with stable, porous gel particles. When the exosome-containing sample solution is loaded onto the SEC column, smaller particles are trapped inside the pores, while larger molecules cannot enter the pores and are eluted first. Accordingly, eluted fractions collected at different stages contain molecules of different sizes: large exosome particles elute first, followed by smaller protein and lipid particles. This kit is well suited for downstream research including exosome functional validation, characterization, and analysis of cargo contents (nucleic acids, proteins, lipids, metabolites detection or omics analysis).
Contents & Storage:
| Component No. | Component Name | Appearance | 1ml | Storage |
| C778164A | SEC Purification Column (1mL) | Liquid | 1 mL | 2-8℃ |
| C778164B | Storage Solution (5×) | Liquid | 30 mL | 2-8℃ |
| C778164C | Washing Solution (5×) | Liquid | 20 mL | 2-8℃ |
Product Features:
High Purity: The removal rate of contaminating proteins reaches up to 99.44%, ensuring the accuracy of subsequent research from the source.
Convenient & Rapid: Simple operation; the whole procedure takes only 30 min.
Stable Results: Excellent reproducibility with highly consistent experimental data.
Product Principle:
Figure 1. Working Principle of Exosome Purification Kit (SEC)
Product Data:
1. NTA Test Results
Figure 2. NTA results show that exosomes are mainly present in fractions 3–7.
2. TEM Test Results
Figure 3. TEM results demonstrate that exosomes purified and collected using SEC Column 2.0 exhibit intact and well-defined morphology.
3. BCA Protein Assay Results
Figure 4. BCA results indicate that the removal rate of contaminating proteins exceeds 99.5%.
Operating Protocol:
Sample Pre-treatment:
1. Cell Culture Supernatant
① Collect 10–50 mL cell culture supernatant (cell density at harvest shall be no less than 1×10⁶ cfu/mL);
② Centrifuge the cell supernatant at 3000 g for 10 min and retain the supernatant;
③ Filter the supernatant sample through a 0.22 μm filter membrane;
④ Concentrate the filtrate with a 100 kDa ultrafiltration tube at a concentration ratio of 1:20. Collect the concentrate for later use.
2. Urine
① Collect approximately 10–50 mL midstream morning urine, centrifuge at 3000 g for 10 min and retain the supernatant;
② Filter the supernatant through a 0.22 μm filter membrane;
③ Concentrate the filtrate with a 100 kDa ultrafiltration tube at a concentration ratio of 1:20. Collect the concentrate for later use.
Exosome Isolation:
1. Reagent Preparation
① Both Storage Solution (5×) and Washing Solution (5×) need to be diluted to 1× with deionized water. All prepared reagents must be filtered through a 0.22 μm filter membrane before use.
② Prepare 1× PBS by yourself. Filter it through a 0.22 μm filter membrane prior to use, or purchase pre-filtered PBS (equilibrate to room temperature before use).
2. Column Equilibration
① Take the SEC purification column out of the refrigerator and fix it vertically (fixing device to be provided by user);
② Let it stand at room temperature for 30 minutes;
③ Place an empty beaker underneath the SEC purification column;
④ Open the top plunger and bottom cap, drain the storage solution inside the column until no liquid flows out from the outlet. Add 15 mL of 1× PBS to the column (can be added in portions). Wait until all liquid flows out before sample loading.
3. Sample Loading
① Prepare 8 EP tubes and label them sequentially as 1, 2, 3, 4, 5, 6, 7, 8;
② Load 1 mL sample slowly along the inner wall from the top of the SEC purification column. After the sample fully enters the packing material and no liquid flows out from the bottom outlet, add 0.5 mL of 1× PBS. Collect the eluate into EP tube 1 with a collection volume of 0.5 mL;
③ After the previous 0.5 mL 1× PBS completely enters the size-exclusion column, add another 0.5 mL of 1× PBS and collect the eluate into EP tube 2 with a collection volume of 0.5 mL.
Note: Keep liquid inside the column during operation to ensure full hydration of the packing material.
④ Repeat step ③ six times to collect a total of 8 tubes of eluate.
Notes: The minimum sample volume is 1 mL. If the sample volume is less than 1 mL, make up to 1 mL with 1× PBS, mix well and then load onto the column.
4. Exosome Collection
① For high exosome recovery: retain fractions 3, 4, 5, 6 and 7, totaling 2.5 mL. If fraction separation is not required, add 2.5 mL of 1× PBS directly after the second 0.5 mL 1× PBS fully enters the size-exclusion column and elute into a single collection tube.
② For high-purity exosomes: combine fractions 4, 5 and 6, totaling 1.5 mL.
Note: If concentration of exosome eluate is required, use a 100 kDa ultrafiltration tube and centrifuge at 4000 g to concentrate to the target volume.
5. Cleaning and Storage of SEC Purification Column
After collection, add 10 mL Washing Solution, followed by 15 mL 1× PBS for flushing, then flush with 10 mL 1× Storage Solution. Close the bottom cap, fill the SEC purification column with 1× Storage Solution, seal the top port with the plunger, verify no leakage, and store at 4 ℃.
Notes: ⁕This SEC purification column can be reused for up to 10 times.
⁕Seal the top of the column with parafilm after use to prevent evaporation of storage solution.
Disclaimer:
This product is for research use only. It shall not be used for clinical diagnosis or therapy, nor for food or pharmaceutical applications.
规格
别名
外泌体纯化试剂盒
英文别名
Exosome Isolation and Purification Kit
规格或纯度
BioReagent, 无菌
英文名称
Cell Culture Media/Urine Exosome Purification Kit (SEC)
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
Store at 2-8℃ long term (12 months).
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 |
|---|---|---|
 ZJ26F0939579 |
分析证书 | C778164 |
| ZJ25F0926930 |
分析证书 | C778164 |
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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