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纯化试剂盒是一款高效、灵活的工具，专门用于从多种样本来源（如血清、腹水、细胞培养上清或杂交瘤上清）中高纯度地纯化抗体。与传统的预装柱试剂盒不同，试剂盒不包含预装柱和填料，您可以根据自己的实验需求，选用合适的填料至任何规格的层析空柱中，完美适配手动操作或自动化纯化系统。 本产品提供高浓度的Protein A/G 纯化buffer，赋予研究人员最大的实验灵活性，可适配于各种层析空柱、重力流柱或批次纯化流程。
产品组份
| P1456508 | 组份 | 10T | 50T | 储存温度 | 单次测试使用量 |
| P1456508A | 结合缓冲液 (10×) | 50 mL | 250 mL | 2-8℃ | 5 mL |
| P1456508B | 洗脱缓冲液 (5×) | 10 mL | 50 mL | 2-8℃ | 1 mL |
| P1456508C | 中和缓冲液 | 10 mL | 50 mL | 2-8℃ | 1 mL |
| P1456508D | 考马斯亮蓝染色液 | 10 mL | 50 mL | 2-8℃ | 1 mL |
| P1455932E | 再生缓冲液 | 10 mL | 50 mL | 2-8℃ | 10 mL (可选) |
注：每个Test推荐纯化填料使用量为2 mL
使用方法
使用前，需用去离子水将各组分缓冲液稀释至 1× 浓度。具体操作可根据不同纯化工具的要求调整。
一．样品制备
在纯化之前，必须通过离心或 0.45 μm 过滤器过滤来澄清杂交瘤组织上清、血清或腹水样品，以免堵塞纯化柱，降低纯化柱寿命。
二．样品纯化
2.1 平衡
取 1× 结合缓冲液（Binding buffer）倒满纯化柱（约 15 个柱体积），用于平衡柱体，使液体自然滴落。
2.2 上样
取下纯化柱上盖，将澄清后的样品加入纯化柱中，使液体自然滴落。
注：若样品中抗体含量较低或样品体积较大，可通过增加上样次数、延长上样时间，提升抗体与填料的结合效率。
2.3 洗涤
取 1× 结合缓冲液（Binding buffer）倒满纯化柱（约 15 个柱体积），清洗柱内未结合的杂质，使液体自然滴落。
2.4 洗脱
1. 准备数个 1.5mL 离心管，向每个离心管中预先加入 10μL 中和缓冲液（Neutralization Buffer）。
2. 向每个纯化柱中加入 3mL 1× 洗脱缓冲液（Elution buffer，约 3 个柱体积），通过重力作用收集洗脱液。每管收集 8 滴洗脱液（约 0.5mL），依次操作直至洗脱完成。
2.5 透析
1. 从每管洗脱液中取 2.5μL，与 10μL 考马斯亮蓝染色液（Coomassie Brilliant Blue Staining Solution）混合，观察抗体洗脱的梯度变化。
2. 收集抗体浓度较高的洗脱管，将其置于适宜的缓冲液中进行透析置换，最终获得高纯度的纯化 IgG。
三．纯化柱储存
3.1 抗体洗脱完成后，继续向柱内加入 2mL 1× 洗脱缓冲液（Elution buffer）洗涤，避免柱内残留抗体。
3.2 取 1× 结合缓冲液（Binding buffer）倒满纯化柱（约 15 个柱体积），平衡柱体，使液体自然滴落。
3.3 取 20% 乙醇倒满纯化柱（约 15 个柱体积），进行防腐处理，使液体自然滴落。
3.4 向柱内加入 5mL 20% 乙醇，盖上上盖并堵紧下方堵头，于 2-8℃条件下储存，下次纯化时可直接取出使用。
四．柱再生 （可选）
当纯化柱的性能出现可观测的下降时，即应执行再生程序，具体操作如下：
4.1 向柱内加入 10mL 再生缓冲液（Regeneration Buffer），进行洗涤。
4.2 加入 30mL 超纯水（ddH₂O）清洗柱体，确保将再生缓冲液彻底冲洗干净。
4.3 加入 20mL 1× 结合缓冲液（Binding buffer）洗涤，平衡填料。
4.4 加入 15mL 20% 乙醇洗涤后，再向柱内加入 5mL 20% 乙醇，盖上上盖并堵紧下方堵头，于 2-8℃条件下储存。
The purification kit is a highly efficient and flexible tool specifically designed for high-purity antibody purification from a variety of sample sources (such as serum, ascites, cell culture supernatant, or hybridoma supernatant). Unlike traditional pre-packed column kits, this kit does not include pre-packed columns or chromatography media. Instead, you can select and pack suitable media into empty columns of any scale according to your experimental needs, making it perfectly adaptable for both manual operation and automated purification systems. This product provides a high-concentration Protein A/G purification buffer, offering researchers maximum experimental flexibility to accommodate various empty chromatography columns, gravity-flow columns, or batch purification processes.
| P1456508 | Components | 10T | 50T | Storage | Quantity Per Test |
| P1456508A | Binding buffer(10×) | 50 mL | 250mL | 2-8℃ | 5 mL |
| P1456508B | Elution buffer(5×) | 10 mL | 50 mL | 2-8℃ | 1 mL |
| P1456508C | Neutralizer Buffer | 10 mL | 50 mL | 2-8℃ | 1 mL |
| P1456508D | Coomassie Brilliant Blue Staining Solution | 10 mL | 50 mL | 2-8℃ | 1 mL |
| P1455932E | Regeneration Buffer | 10 mL | 50 mL | 2-8℃ | 10mL (Optional) |
Note: The recommended amount of Purification resin for each test is 2 mL.
Instructions for Use
Prior to use, dilute all component buffers to 1× concentration with deionized water. Specific operations can be adjusted according to the requirements of different purification tools.
I. Sample Preparation
Before purification, clarify hybridoma supernatants, serum, or ascites samples by centrifugation or filtration through a 0.45 μm filter to avoid clogging the purification column and reducing its lifespan.
II. Sample Purification
1. Equilibration
Fill the purification column with 1× Binding Buffer (approximately 15 column volumes) to equilibrate the column, allowing the liquid to drain naturally.
2. Sample Loading
Remove the top cap of the purification column and add the clarified sample to the Protein A purification column, allowing the liquid to drain naturally.
Note: If the antibody concentration in the sample is low or the sample volume is large, increase the number of loading cycles or extend the loading time to improve the binding efficiency between the antibody and the resin.
3. Washing
Fill the purification column with 1× Binding Buffer (approximately 15 column volumes) to wash away unbound impurities, allowing the liquid to drain naturally.
4. Elution
4.1 Prepare several 1.5 mL centrifuge tubes and pre-add 10 μL of Neutralizer Buffer to each tube.
4.2 Add 3 mL of 1× Elution Buffer (approximately 3 column volumes) to each purification column and collect the eluate by gravity. Collect 8 drops (approximately 0.5 mL) per tube and repeat until elution is complete.
5. Dialysis
5.1 Take 2.5 μL of eluate from each tube, mix with 10 μL of Coomassie Brilliant Blue Staining Solution, and observe the gradient change of antibody elution.
5.2 Collect the eluate tubes with high antibody concentration and dialyze against an appropriate buffer to obtain highly purified IgG.
III. Storage of Purification Column
1. After antibody elution, add an additional 2 mL of 1× Elution Buffer to the column to wash away any residual antibody.
2. Fill the purification column with 1× Binding Buffer (approximately 15 column volumes) to equilibrate the column, allowing the liquid to drain naturally.
3. Fill the purification column with 20% ethanol (approximately 15 column volumes) for preservation, allowing the liquid to drain naturally.
4. Add 5 mL of 20% ethanol to the column, cap the top, and seal the bottom cap. Store at 2-8℃ for direct use in subsequent purifications.
IV. Column Regeneration (Optional)
Perform regeneration when a measurable decrease in column performance is observed, following these steps:
1. Add 10 mL of Regeneration Buffer to the column for washing.
2. Add 30 mL of double-distilled water (ddH₂O) to rinse the column thoroughly and remove residual Regeneration Buffer.
3. Add 20 mL of 1× Binding Buffer to wash and equilibrate the resin.
4. Add 15 mL of 20% ethanol for washing, then add 5 mL of 20% ethanol to the column. Cap the top and seal the bottom cap, and store at 2-8℃
| P1456508 | 组份 | 10T | 50T | 储存温度 | 单次测试使用量 |
| P1456508A | 结合缓冲液 (10×) | 50 mL | 250 mL | 2-8℃ | 5 mL |
| P1456508B | 洗脱缓冲液 (5×) | 10 mL | 50 mL | 2-8℃ | 1 mL |
| P1456508C | 中和缓冲液 | 10 mL | 50 mL | 2-8℃ | 1 mL |
| P1456508D | 考马斯亮蓝染色液 | 10 mL | 50 mL | 2-8℃ | 1 mL |
| P1455932E | 再生缓冲液 | 10 mL | 50 mL | 2-8℃ | 10 mL (可选) |
Note: The recommended amount of Purification resin for each test is 2 mL.
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 |
|---|---|---|
 ZJ26F0535821 |
分析证书 | P1456508 |
危险品化学品经营许可证（带存储）