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1 μm MPSE-SAX 固相萃取磁珠（强阴离子交换）由二氧化硅、聚苯乙烯和纳米氧化铁组成，能够快速从复杂生物样品（例如血清、血浆、尿液）中分离蛋白质和肽。本产品可结合样品中的弱阳离子，这些弱阳离子通常不能与弱阴离子交换磁珠。固相萃取磁珠结合了固相萃取技术和磁性分离技术，可广泛应用于样品前处理，在环境监测污染物检测、药物分析中的代谢物提取、食品安全检测、毒素或残留农药提取等方面广泛应用。磁珠表面通过功能化处理修饰有特定的吸附官能团，可通过疏水、静电或特异性结合作用力选择性吸附目标物，样品准备快速简单，经外加磁场可方便快捷地分离，从而提高样品前处理的效率和效果。
| 货号 | M1520788 |
| 产品名称 | 1 μm MPSE-SAX 固相萃取磁珠 |
| 浓度 | 10 mg/mL |
| 粒径 | 约1 μm |
| 表面电位 | -20 mV左右 |
| 磁含量 | 大约35% - 45% |
| 分散溶剂 | 纯水 |
使用说明
注意：从技术上来说，很难设计出一种通用的采用 1 μm MPSE-SAX（强阴离子交换）固相萃取磁珠进行蛋白纯化的方案。以下方案是采用1 μm MPSE-SAX（强阴离子交换）固相萃取磁珠对蛋白质和/或肽混合物进行分级纯化的示例。为了达到最佳的使用效果，用户可以根据此方案采用替代的结合、洗涤和洗脱缓冲液，同时通过滴定法获得每种应用中最佳的磁珠用量以及最佳的洗脱体积。
1. 选择合适的离子交换磁珠
带电的蛋白质或肽会与相反电荷基质结合。如果目标蛋白质或肽的等电点（pI）高于缓冲溶液的 pH 值，则蛋白质或肽表面带有正电荷。需选择使用阴离子交换磁珠MPSE-WAX（弱阴离子交换）或MPSE-SAX（强阴离子交换）磁珠进行纯化。如果目标蛋白质或肽的等电点低于缓冲溶液的 pH 值，则蛋白质或肽表面带有负电荷，则应选择阳离子交换磁珠1 μm MPSE-WCX（弱阳离子交换）或MPSE-SCX（强阳离子交换）磁珠进行纯化。
2. 选择合适的缓冲溶液
结合/ 洗涤缓冲液：结合/洗涤缓冲液的 pH 值应与蛋白质或肽的等电点至少相差一个 pH 单位。
洗脱缓冲溶液：要从磁珠上洗脱目标蛋白或肽，用户应针对具体的应用逐步优化洗脱条件。可采用盐浓度逐步升高的溶液（0.1-1.0 M NaCl），使用 0.5%三氟乙酸（TFA），或改变洗脱缓冲溶液的 pH（例如，20 mMTris pH 7.0，随后是 20 mM MES pH 6.0，随后是 20 mM MES pH 5.0，50 mM 柠檬酸缓冲液 pH 4.0）进行逐步洗脱。
参考实验步骤
预处理缓冲液：含有 1M NaCl 的 50 mM PB 溶液，pH8.0。
结合/ 洗涤缓冲液：含有 20 mM NaCl 的 50 mM PB 溶液，pH 8.0。
A. 磁珠准备
1. 充分分散磁珠溶液，取100 μL磁珠（10 mg/mL）磁珠于离心管中，将离心管置于磁分离器上 1-3 分钟使磁珠充分磁分离，去除上清。
2. 取 200 μL 预处理缓冲液将磁珠充分重悬，将离心管置于室温下平衡 2-3 分钟。将离心管置于磁分离器上 1-3 分钟，使磁珠充分磁分离，去除上清。
3. 重复步骤 2 一次。
4. 从磁分离器上取下离心管，用 200 μL 结合/洗涤缓冲液将磁珠充分重悬。将离心管置于磁分离器上 1-3分钟，使磁珠充分磁分离，去除上清液。
5. 重复步骤 4 一次。
6. 用结合/洗涤缓冲液将磁珠充分重悬。
B. 蛋白结合
1. 将含有 60 μg 的蛋白质或肽的样品（注意：为获得最佳效果，应将样品悬浮于结合/洗涤缓冲液中）加入到步骤 A-6 中已洗涤过的磁珠所在的离心管中。如有必要，用结合/洗涤缓冲液将总体积调整至 200 μL。用移液器充分混匀磁珠，在室温下放置 2-3 分钟。
2. 将离心管置于磁分离器上 1-3 分钟，使磁珠充分磁分离，吸去上清液。
3. 从磁分离器上取下离心管，用 200 μL 结合/洗涤缓冲液清洗磁珠。将离心管置于磁分离器上 1-3 分钟，充分磁分离，去除上清液。
4. 重复步骤 3 三次。
C. 洗脱
1. 从磁分离器上取下离心管，用 200 μL 结合/洗涤缓冲液重悬磁珠，在室温放置 3 分钟。
2. 将离心管置于磁分离器上 1-3 分钟，充分磁分离，然后将含有洗脱蛋白的上清液转移至新的离心管中，进行下游检测。
常见问题与解答
| 问题 | 可能的原因 | 建议 |
| 低产率 | 离子相互作用不够强 结合/洗涤缓冲溶液的 pH 值高于目标蛋白或肽的等电点 | 优化结合/洗涤缓冲溶液中的氯化钠浓度 优化结合缓冲液的 pH 值 |
| 洗脱不良 | 离子相互作用太强了 | 优化氯化钠浓度 优化洗脱缓冲溶液的 pH 值 使用弱阴离子交换磁珠 |
| 纯化效果不佳 | 洗涤不充分 | 在每次洗脱步骤之前增加洗涤步骤 |
| 纯化效果不佳 | 可能存在与目标蛋白质或肽等电点相似的蛋白质或肽 | 优化洗脱缓冲溶液的氯化钠浓度或 pH 值 |
注意事项
磁珠取用前应充分混匀，防止取用改变磁珠浓度，避免长时间超声对磁珠表面破坏。
1 μm MPSE-SAX Solid Phase Extraction Magnetic Beads (Strong Anion Exchange) are composed of silica, polystyrene, and iron oxide nanoparticles, enabling rapid separation of proteins and peptides from complex biological samples (e.g., serum, plasma, urine). This product can bind weakly cationic species in samples that typically cannot be captured by weak anion exchange magnetic beads. Solid phase extraction magnetic beads combine solid phase extraction technology with magnetic separation technology and are widely used in sample pretreatment, with applications in environmental monitoring for pollutant detection, metabolite extraction in pharmaceutical analysis, food safety testing, and extraction of toxins or pesticide residues, among others. The bead surface is functionalized with specific functional groups, allowing selective adsorption of targets through hydrophobic, electrostatic, or specific interactions. Sample preparation is rapid and straightforward, and the beads can be conveniently and quickly separated using an external magnetic field, thereby improving the efficiency and effectiveness of sample pretreatment.
| Cat. No. | M1520788 |
| Product Name | 1 μm MPSE-SAX Solid Phase Extraction Magnetic Beads |
| Concentration | 10 mg/mL |
| Particle Size | Approx. 1 μm |
| Surface Potential | Approx. -20 mV |
| Magnetic Content | Approx. 35% – 45% |
| Dispersion Solvent | Pure Water |
Instructions for Use
Note: Technically, it is challenging to design a universal protocol for protein purification using 1 μm MPSE-SAX (Strong Anion Exchange) Solid Phase Extraction Magnetic Beads. The following protocol is an example of using 1 μm MPSE-SAX (Strong Anion Exchange) Solid Phase Extraction Magnetic Beads for the fractionation and purification of protein and/or peptide mixtures. For optimal results, users may adapt alternative binding, washing, and elution buffers based on this protocol, while determining the optimal bead quantity and elution volume for each application through titration experiments.
1. Selecting the Appropriate Ion Exchange Magnetic Beads
Charged proteins or peptides bind to matrices of the opposite charge. If the isoelectric point (pI) of the target protein or peptide is higher than the pH of the buffer solution, the protein or peptide surface carries a positive charge. In this case, anion exchange magnetic beads, MPSE-WAX (Weak Anion Exchange) or MPSE-SAX (Strong Anion Exchange), should be selected for purification. If the isoelectric point of the target protein or peptide is lower than the pH of the buffer solution, the protein or peptide surface carries a negative charge, and cation exchange magnetic beads, such as 1 μm MPSE-WCX (Weak Cation Exchange) or MPSE-SCX (Strong Cation Exchange), should be selected for purification.
2. Selecting the Appropriate Buffer Solutions
Binding/Washing Buffer: The pH of the binding/washing buffer should differ from the isoelectric point of the protein or peptide by at least one pH unit.
Elution Buffer: To elute the target protein or peptide from the beads, users should gradually optimize the elution conditions for their specific application. Elution can be performed using solutions with increasing salt concentrations (0.1–1.0 M NaCl), using 0.5% trifluoroacetic acid (TFA), or by changing the pH of the elution buffer (e.g., 20 mM Tris pH 7.0, followed by 20 mM MES pH 6.0, followed by 20 mM MES pH 5.0, 50 mM citrate buffer pH 4.0) for stepwise elution.
Reference Experimental Procedure
Pre-treatment Buffer: 50 mM PB solution containing 1 M NaCl, pH 8.0.
Binding/Washing Buffer: 50 mM PB solution containing 20 mM NaCl, pH 8.0.
A. Bead Preparation
Thoroughly disperse the bead solution. Transfer 100 μL of magnetic beads (10 mg/mL) into a centrifuge tube. Place the tube on a magnetic separator for 1–3 minutes to allow complete magnetic separation, then remove the supernatant.
Add 200 μL of pre-treatment buffer to fully resuspend the beads. Allow the tube to equilibrate at room temperature for 2–3 minutes. Place the tube on the magnetic separator for 1–3 minutes for complete magnetic separation, then remove the supernatant.
Repeat step 2 once.
Remove the tube from the magnetic separator. Resuspend the beads thoroughly in 200 μL of binding/washing buffer. Place the tube on the magnetic separator for 1–3 minutes for complete magnetic separation, then remove the supernatant.
Repeat step 4 once.
Resuspend the beads thoroughly in binding/washing buffer.
B. Protein Binding
Add the sample containing 60 μg of protein or peptide (Note: For optimal results, the sample should be suspended in binding/washing buffer) to the centrifuge tube containing the washed beads from Step A-6. If necessary, adjust the total volume to 200 μL with binding/washing buffer. Mix the beads thoroughly by pipetting and incubate at room temperature for 2–3 minutes.
Place the centrifuge tube on the magnetic separator for 1–3 minutes for complete magnetic separation, then aspirate and discard the supernatant.
Remove the tube from the magnetic separator. Wash the beads with 200 μL of binding/washing buffer. Place the tube on the magnetic separator for 1–3 minutes for complete magnetic separation, then remove the supernatant.
Repeat step 3 three times.
C. Elution
Remove the tube from the magnetic separator. Resuspend the beads in 200 μL of binding/washing buffer and incubate at room temperature for 3 minutes.
Place the centrifuge tube on the magnetic separator for 1–3 minutes for complete magnetic separation. Transfer the supernatant containing the eluted protein to a new centrifuge tube for downstream analysis.
Frequently Asked Questions and Troubleshooting
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Precautions
Ensure the magnetic beads are thoroughly mixed before use to prevent changes in concentration during pipetting. Avoid prolonged ultrasonication, which may damage the bead surface.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0535082 |
分析证书 | M1520788 |
| ZJ26F0434421 |
分析证书 | M1520788 |
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