BioReagent 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
2-8°C储存,禁止冷冻。低温运输,禁止冷冻 。请查阅批次 COA 获取详细规格。
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。
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。
储存缓冲液：100% Isopropyl Alcohol
UltraBio™ 环氧基磁珠 (1μm) 是一种环氧活化的具有超顺磁性的磁性微粒，是一种被广泛应用的功能性生物磁珠，主要用于生物分析检测，可以直接用于含氨基、巯基和羟基的蛋白和样品的耦联。预活化介质可以根据需要制备成特殊的亲和介质，快速有效地从复杂体系中一步纯化相应的物质。
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使用说明：
一、缓冲液准备
偶联液：0.1M Na2CO3，pH 9.5
封闭液：1M 乙醇胺，pH 8.0
清洗液1：0.1M 乙酸-乙酸钠，0.5M NaCl，pH 3.0
清洗液2：0.1M Tris-HCl，0.5M NaCl，pH 8.0
保护液：PBS，0.01% Tween-20，0.02% NaN3
注：样品用偶联液溶解，浓度约5-10 mg/mL。偶联液可以选择碳酸盐、磷酸盐等不含氨基的缓冲液体系。
二、样品偶联
1、准备适量的UltraBio™ 环氧基磁珠 (1μm)（磁珠浓度为10mg/mL），将磁珠反复颠倒，充分混匀，使用移液器取适量的磁珠悬浮液，置于离心管中，将离心管置于磁分离器上，大约1min，待溶液变澄清后，用移液器吸弃清液。
2、再将离心管磁分离器上取下来，加入去离子水，使用枪头反复吹打5-10次，将离心管置于磁分离器上，大约1min，待溶液变澄清后，用移液器吸弃清液，重复洗涤1次。再用偶联液清洗1次。 3、将准备好的样品加入到处理好的磁珠中（磁珠在样品中浓度约30-50mg/ml），颠倒混匀。将离心管置于混合仪上，室温孵育24至48h。确保磁珠混匀，否则影响偶联效率。
4、将离心管置于磁分离器，大约1min，待溶液变澄清后，用移液器移出上清液，保留，以备取样检测。分别用去离子水清洗，清洗液1、去离子水、清洗液2和去离子水重复清洗磁珠2次，然后保存在等体积的保护液中，于2-8℃保存。
三、纯化流程
下面以磁珠偶联抗体后，纯化抗原为例，介绍纯化步骤。
1、缓冲液的准备
所用水和缓冲液在使用之前建议用0.22μm或0.45μm滤膜过滤。
平衡/洗杂液：0.15M NaCl，20mM Na2HPO4，pH 7.0
洗脱液：0.1M 甘氨酸，pH 3.0
中和液：1M Tris-HCl，pH 8.5
2、磁珠预处理
将偶联好的磁珠混合均匀，取计算量（根据上样量和磁珠载量计算）的磁珠悬浮液（后文均以100μl为例），转移至离心管中，放置在磁分离器上，静置大约1min，待溶液变澄清后，用移液器吸弃上清液。再将离心管从磁分离器上取下来，加入与所取磁珠悬浮液等体积（100 μl）的平衡液，使用枪头反复吹打5次，将离心管置于磁分离器上，大约1min，待溶液变澄清后，用移液器吸弃上清液，重复洗涤2次。
3、样品吸附
在步骤2预处理的磁珠中加入样品溶液，漩涡振荡均匀，在室温下置于翻转混合仪或者手工轻轻翻转离心管，促使样品和磁珠充分接触并吸附，混合30min以上（具体时间根据结合效果调整），置于磁分离器上，大约1min，待溶液变澄清后，吸弃上清液。
4、洗杂
向离心管中加入初始磁珠悬浮液5倍体积（500μl）的洗杂液，振荡悬浮，混合1-2min，然后置于磁分离器上，大约1min，待溶液变澄清后，吸弃上清液。该操作重复2次。
5、洗脱
在上述离心管中加入3-5倍悬浮液体积（300-500μl）的洗脱液，用移液器吹打5次， 然后在室温下置于翻转混合仪或者手工轻轻翻转离心管，5-10min后，置于磁分离器上，大约1min，待溶液变澄清后，吸取上清液，收集洗脱组分，即为目标蛋白。
6、洗脱组分中和
向洗脱组分中加入洗脱体积十分之一的中和液，调节pH值至7.0-8.0。
7、磁珠保存
使用后的磁珠用1ml洗脱液重悬磁珠，然后置于磁分离器上，大约1min，待溶液变澄清后，吸弃上清液。该操作重复2次。再加入1ml 平衡液，悬浮磁珠，然后置于磁分离器上，大约1min，待溶液变澄清后，吸弃上清液。加入步骤一中的保护液至磁珠浓度为10mg/ml， 置于2-8℃保存。
UltraBio™ Epoxy Magnetic Beads (1 μm) are epoxy-activated, superparamagnetic magnetic particles. As a widely utilized functional biological magnetic bead, they are primarily used for bioanalytical detection. These beads enable direct coupling with proteins and samples containing amino, thiol, or hydroxyl groups. This pre-activated medium can be customized to prepare specialized affinity media for the rapid and efficient one-step purification of target substances from complex systems.
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Instructions for Use:
I. Buffer Preparation
Coupling Buffer: 0.1 M Na₂CO₃, pH 9.5
Blocking Solution: 1 M Ethanolamine, pH 8.0
Wash Buffer 1: 0.1 M Sodium Acetate-Acetic Acid, 0.5 M NaCl, pH 3.0
Wash Buffer 2: 0.1 M Tris-HCl, 0.5 M NaCl, pH 8.0
Storage Solution: PBS, 0.01% Tween-20, 0.02% NaN₃
*Note: Dissolve the sample in Coupling Buffer to a concentration of ~5-10 mg/mL. Coupling buffers (e.g., carbonate, phosphate) should lack primary amine groups.*
II. Sample Coupling
Invert the UltraBio™ Epoxy-Activated Magnetic Beads (1 μm) vial repeatedly to ensure thorough mixing (bead concentration: 10 mg/mL). Transfer the desired volume of bead suspension to a microcentrifuge tube.
Place the tube on a magnetic rack for ~1 minute until the solution clears. Aspirate and discard the supernatant.
Remove the tube from the rack. Add deionized water. Pipette mix 5-10 times to resuspend the beads.
Place the tube on the magnetic rack for ~1 minute. Aspirate and discard the supernatant.
Repeat Steps 3 & 4 once more (2 water washes total).
Wash the beads once with Coupling Buffer (Steps 3 & 4).
Add the prepared sample solution to the washed beads. The final bead concentration in the sample mixture should be ~30-50 mg/mL. Mix by inversion.
Incubate the tube with end-over-end rotation at room temperature for 24-48 hours. Ensure the beads remain uniformly suspended during incubation to maximize coupling efficiency.
Place the tube on the magnetic rack for ~1 minute. Aspirate and save the supernatant for coupling efficiency analysis if needed.
Wash the beads sequentially with:
Deionized Water
Wash Buffer 1
Deionized Water
Wash Buffer 2
Deionized Water
Perform magnetic separation and aspirate supernatant after each wash. Repeat this alternating wash cycle once more (2 cycles total).
Resuspend the coupled beads in an equal volume of Storage Solution. Store at 2-8°C.
III. Purification Procedure (Example: Antigen Purification using Antibody-Coupled Beads)
Buffer Preparation:
Filter all water and buffers (0.22 μm or 0.45 μm) before use.
Equilibration/Wash Buffer: 0.15 M NaCl, 20 mM Na₂HPO₄, pH 7.0
Elution Buffer: 0.1 M Glycine, pH 3.0
Neutralization Buffer: 1 M Tris-HCl, pH 8.5
Bead Pre-equilibration:
Resuspend coupled beads thoroughly. Transfer the calculated volume (e.g., 100 μL) to a microcentrifuge tube.
Place on magnetic rack for ~1 min until solution clears. Aspirate supernatant.
Remove tube from rack. Add an equal volume (e.g., 100 μL) of Equilibration Buffer. Pipette mix 5 times.
Place on magnetic rack for ~1 min. Aspirate supernatant.
Repeat equilibration wash twice (3 washes total).
Sample Binding:
Add sample solution to the pre-equilibrated beads. Vortex-mix briefly.
Incubate with end-over-end rotation (or gentle manual tube inversion) at room temperature for ≥30 min (adjust based on binding efficiency).
Place on magnetic rack for ~1 min. Aspirate and discard supernatant.
Washing:
Add 5x initial bead volume (e.g., 500 μL) of Wash Buffer. Vortex-mix to resuspend.
Incubate with mixing for 1-2 min.
Place on magnetic rack for ~1 min. Aspirate and discard supernatant.
Repeat wash twice (3 washes total).
Elution:
Add 3-5x initial bead volume (e.g., 300-500 μL) of Elution Buffer. Pipette mix 5 times.
Incubate with end-over-end rotation (or gentle manual tube inversion) at room temperature for 5-10 min.
Place on magnetic rack for ~1 min. Transfer the clear supernatant (elution fraction) to a clean tube. This contains the target protein.
Neutralization:
Immediately add 1/10th volume of the elution fraction of Neutralization Buffer to the elution fraction. Mix thoroughly to adjust pH to 7.0-8.0.
Bead Regeneration & Storage:
Add 1 mL Elution Buffer to the beads. Vortex-mix. Place on magnetic rack for ~1 min. Aspirate supernatant.
Repeat elution wash once (2 washes total).
Add 1 mL Equilibration Buffer. Vortex-mix. Place on magnetic rack for ~1 min. Aspirate supernatant.
Resuspend beads in Storage Solution (Section I) at 10 mg/mL. Store at 2-8°C.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F1039791 |
分析证书 | E1456039 |
| ZJ26F0535713 |
分析证书 | E1456039 |
| ZJ26F0535712 |
分析证书 | E1456039 |
| ZJ26F0231923 |
分析证书 | E1456039 |
| ZJ25F0826357 |
分析证书 | E1456039 |
| ZJ25F0826356 |
分析证书 | E1456039 |
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