DEAE琼脂糖磁珠是一种弱阴离子交换磁珠，具有快速的磁响应性、丰富的离子交换能力和极高的蛋白结合能力。离子交换配基为二乙氨乙基（diethylaminoethyl，DEAE），该配基在 pH 3-12 的工作范围内仍然能够保持稳定的高蛋白结合能力。与传统柱层析纯化方式相比，Magrose DEAE 磁珠无需对粗蛋白样品进行预处理（如：反复繁琐的离心，费时费力的过滤操作），此外，无需控制流速及柱压，不需要昂贵的层析设备。对于熟练的操作者来说，在很短时间内就能完成高纯度目的蛋白的提取，且能轻松实现多个样品的平行处理，实现高通量的蛋白纯化。
本产品每毫升磁珠悬浊液含约10% v/v琼脂糖磁珠沉淀。
包装规格： 10% v/v悬液
参数 | 指标 | 磁珠粒径范围 | 30-150 μm | 离子交换类型 | 弱阴离子基团 | 总离子能力 | 110~170 μmol/mL Gel | 蛋白结合量 ① | ≥110 mg BSA/mL Gel | 保存液 | 20% 乙醇 | 悬液浓度 ② | 10%(V/V) 磁珠悬液 | 保存温度 | 2-8℃ | 工作 pH 范围 | 3-12 | 保质期 | 2年 |
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注：
①：磁珠蛋白结合量与目标蛋白特性有关，此处仅做参考值
②：1 mL 磁珠悬液中包含 100 μL 磁珠
产品优势
1. 磁响应速度快，减少操作时间
2. 磁珠具有良好的分散性和重悬性，提高操作的便捷性
3. 配基具有良好的物理化学稳定性，提高了实验结果的可靠性及可重复性
操作流程 （以纯化含 BSA 蛋白样品为例）
1. 磁珠预处理（平衡）
将DEAE琼脂糖磁珠漩涡振荡 30 s，使磁珠充分重悬；取一定量的 10%（V/V）磁珠悬液置于 50 mL 离心管中。对磁珠悬液进行磁性分离，弃上清液，加入 20 mL 平衡缓冲液洗涤磁珠 3 次，每次垂直混合 2 min。
注：为了获得目标蛋白的最大回收率，实验者需要加入过量DEAE琼脂糖磁珠，一般大于蛋白结合量的 20%即可。对于含较低丰度目标蛋白的样品中，磁珠对目标蛋白的回收率会有所降低，因此需要继续增大磁珠的用量。
2. 蛋白吸附
将预处理的磁珠加入含 BSA 蛋白的样品溶液中，漩涡振荡 30 s，置于垂直混合仪混匀30~60 min，使样品和磁珠充分接触并吸附，然后进行磁性分离，移弃上清液。
注：为了更有效率的吸附结合物质，平衡缓冲液最好含有较低的离子强度，选择的 pH 值应该至少与目标蛋白的等电点相差一个 pH 单位，选定的盐缓冲液 pH 波动在 0.5 pH 以内。目标蛋白与磁珠的吸附时间与蛋白的本身属性有关。
3. 磁珠洗涤
加入 20 mL 的平衡缓冲液，漩涡振荡重悬磁珠 30 s 后进行磁性分离，移弃上清液；该操作重复 3 次。
4. 蛋白洗脱
在上述完成磁珠洗涤的离心管中加入适量的洗脱液进行蛋白洗脱。用移液器吹打或者涡旋振荡下迅速重悬，然后将离心管置于垂直混合仪混匀 10~15 min 后进行磁性分离，收集上清液至新的离心管中。洗脱方式主要有高盐浓度洗脱（含 1-2 M 的 NaCl 的平衡缓冲液）和低 pH 洗脱（选择低于目标蛋白等电点的 pH范围即可）两种。
5. 磁珠再生
一般采用 2 M NaCl 溶液洗涤 3~5 次，然后用平衡缓冲液进行再平衡处理。磁珠多次使用后会有沉淀蛋白、强疏水性蛋白、脂蛋白等杂质非特异性吸附到磁珠上，为了保证磁珠的使用效率，建议进行在位清洗（CIP）。
6. 在位清洗（CIP）
依次采用 1.0 M NaOH、70%乙醇或 30%异丙醇、纯化水洗涤磁珠 2 次；最后加入20%乙醇重悬磁珠，置于 2~8℃保存。
注意事项
1. 本产品不宜冷冻、干燥或离心操作。冷冻、干燥和离心等操作会引起磁珠团聚，不易于重悬和分散， 并且影
响磁珠表面功能基团的化学活性；
2. 在使用本产品前，请务必充分振荡或超声使磁珠保持均匀的悬浮状态；
3. 使用过程中可根据需求，用纯化水或缓冲液磁吸洗涤磁珠 2~3 次，以去除保存液中乙醇；
4. 本产品需与磁性分离设备配套使用；
5. 盐浓度与 pH 值均会影响特异性蛋白的结合与洗脱，客户需自行摸索不同蛋白的结合和洗脱条件，以保证蛋白纯化量和纯度；
6. 本产品仅供研究使用。
DEAE Agarose Magnetic Beads are weak anion-exchange magnetic beads featuring rapid magnetic responsiveness, high ion-exchange capacity, and extremely high protein-binding capacity. The ion-exchange ligand is diethylaminoethyl (DEAE), which maintains stable high protein-binding capacity across a working pH range of 3–12. Compared with traditional column chromatography purification, Magrose DEAE beads eliminate the need for pretreatment of crude protein samples (e.g., repeated and cumbersome centrifugation, time-consuming and laborious filtration). In addition, there is no need to control flow rate or column pressure, and no expensive chromatography equipment is required. For skilled operators, highly pure target proteins can be extracted within a very short time, and parallel processing of multiple samples can be easily achieved for high-throughput protein purification.
Each milliliter of this product's bead suspension contains approximately 10% (v/v) agarose magnetic bead sediment.
Package Specification: 10% (v/v) suspension.
| Parameter | Specification |
|---|
| Bead size range | 30–150 μm | | Ion-exchange type | Weak anion-exchange group | | Total ion capacity | 110–170 μmol/mL Gel | | Protein binding capacity ① | ≥110 mg BSA/mL Gel | | Storage solution | 20% ethanol | | Suspension concentration ② | 10% (V/V) bead suspension | | Storage temperature | 2–8°C | | Working pH range | 3–12 | | Shelf life | 2 years |
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Notes:
① The protein-binding capacity of the beads is related to the characteristics of the target protein; this value is provided for reference only.
② 1 mL of bead suspension contains 100 μL of settled beads.
Product Advantages
1.Fast magnetic response, reducing operation time.
2.Excellent dispersibility and resuspendability of the beads, improving operational convenience.
3.High physicochemical stability of the ligand, enhancing the reliability and reproducibility of experimental results.
Operating Procedure (using purification of BSA-containing protein samples as an example)
1. Bead Pre-treatment (Equilibration)
Vortex the DEAE Agarose Magnetic Beads for 30 s to fully resuspend the beads. Transfer a certain volume of the 10% (V/V) bead suspension into a 50 mL centrifuge tube. Magnetically separate the beads, discard the supernatant, and wash the beads three times with 20 mL of equilibration buffer each time, with vertical mixing for 2 min per wash.
Note: To maximize recovery of the target protein, the user should add an excess of DEAE Agarose Magnetic Beads—generally at least 20% more than the protein-binding capacity. For samples containing lower-abundance target proteins, the recovery rate may decrease; therefore, the bead amount should be further increased.
2. Protein Adsorption
Add the pre-treated beads to the sample solution containing the BSA protein. Vortex for 30 s, then mix on a vertical mixer for 30–60 min to ensure full contact and adsorption between the sample and the beads. Subsequently, perform magnetic separation and discard the supernatant.
Note: For more efficient adsorption of the target substance, the equilibration buffer should preferably have a low ionic strength. The selected pH should be at least one pH unit away from the isoelectric point (pI) of the target protein, and the pH of the chosen salt buffer should fluctuate within 0.5 pH units. The adsorption time between the target protein and the beads depends on the intrinsic properties of the protein.
3. Bead Washing
Add 20 mL of equilibration buffer, vortex for 30 s to resuspend the beads, perform magnetic separation, and discard the supernatant. Repeat this operation three times.
4. Protein Elution
Add an appropriate volume of elution buffer to the centrifuge tube containing the washed beads to elute the protein. Rapidly resuspend the beads by pipetting or vortexing, then place the tube on a vertical mixer for 10–15 min. After magnetic separation, collect the supernatant into a new centrifuge tube. Two main elution methods are available: high-salt elution (equilibration buffer containing 1–2 M NaCl) and low-pH elution (using a pH below the pI of the target protein).
5. Bead Regeneration
Generally, wash the beads 3–5 times with 2 M NaCl solution, followed by re-equilibration with equilibration buffer. After multiple uses, precipitated proteins, strongly hydrophobic proteins, lipoproteins, and other impurities may adsorb nonspecifically to the beads. To ensure efficient bead usage, it is recommended to perform clean-in-place (CIP).
6. Clean-in-Place (CIP)
Sequentially wash the beads twice with 1.0 M NaOH, 70% ethanol or 30% isopropanol, and purified water. Finally, resuspend the beads in 20% ethanol and store at 2–8°C.
Precautions
1.This product should not be frozen, dried, or centrifuged. Freezing, drying, and centrifugation can cause bead aggregation, making resuspension and dispersion difficult, and may affect the chemical activity of the surface functional groups.
2.Before using this product, be sure to vortex or sonicate thoroughly to maintain a uniform suspension of the beads.
3.During use, the beads may be washed 2–3 times with purified water or buffer under magnetic separation to remove the ethanol in the storage solution as needed.
4.This product must be used with magnetic separation equipment.
5.Both salt concentration and pH affect the binding and elution of specific proteins. Users should determine the optimal binding and elution conditions for different proteins to ensure protein yield and purity.
6.This product is for research use only.
危险品化学品经营许可证（带存储）