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：Dl water, 0.05% KroVin300.
UltraBio™ 羧基磁珠 (1μm) 是一种高饱和磁化强度的磁性微球，其表面修饰羧基功能团。UltraBio™ 羧基磁珠 (1μm) 表面的羧基功能团能够在特殊化学试剂（如EDC）的作用下将多肽、蛋白、抗体、寡聚核苷酸等生物配体共价偶联到微球表面，从而可以快速地将目标物从样品中分类和富集，是医学与分子生物学研究中重要的载体工具。
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使用说明：
一、缓冲液准备
活化溶液：50mM MES，pH 5.0
EDC溶液：10mg/ml，50mM MES，pH 5.0 现配现用
封闭液：50mM Tris，pH7.4或50mM 乙醇胺，pH 8.0
清洗液：0.1% Tween-20或Triton X-100的1×PBS溶液，pH 7.4
保护液：Dl water, 0.05% kv300
二、磁珠活化
1、取200µl UltraBio™ 羧基磁珠 (1μm) 于离心管中，在样品混合仪上振荡，充分混匀。
2、将离心管置于磁分离器上约1min，待溶液变澄清后，用移液器吸弃清液。
注：不要吸掉磁珠，下同。
3、将离心管磁分离器上取下来，加入与悬浮液等体积的活化溶液，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。
4、加入200µl（10mg/ml）EDC溶液，混合均匀。
5、室温孵育30min。确保磁珠充分混匀，否则影响活化效率。
6、将离心管置于磁分离器上约1min，待溶液变澄清后，用移液器吸弃清液。用预冷的去离子水和活化溶液清洗2次，尽可能的快速，避免活化基团的水解。
三、配体偶联
1、清洗好的活化磁珠中加入200µl含1-5mg/ml 抗体蛋白的活化溶液，充分混匀，室温孵育过夜, 确保磁珠充分混匀。
2、将离心管置于磁分离器上约1min，待溶液变澄清后，用移液器吸弃清液。加入500µl封闭液，室温孵育30min。
3、可以选择清洗液清洗2次去除非特异性吸附，1×PBS溶液清洗2次，待用。如果暂时不用需要长期保存，可将偶联好配体的磁珠保存在保护液中，2-8℃保存。
四、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，pH7.0
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。该操作重复两次。
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1/10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。该操作重复两次。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™ Carboxyl Magnetic Beads (1 μm) are magnetic microspheres with high saturation magnetization, surface-functionalized with carboxyl groups. The carboxyl groups on these beads enable covalent coupling of biological ligands (such as peptides, proteins, antibodies, and oligonucleotides) to the microsphere surface via specific chemical reagents (e.g., EDC). This facilitates rapid isolation and enrichment of target molecules from complex samples, making them a vital tool in medical and molecular biology research.
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Instructions for Use:
I. Buffer Preparation
Activation Solution: 50 mM MES, pH 5.0
EDC Solution: 10 mg/mL in 50 mM MES, pH 5.0 (Prepare fresh)
Blocking Solution: 50 mM Tris, pH 7.4 OR 50 mM Ethanolamine, pH 8.0
Wash Solution: 1× PBS, pH 7.4 containing 0.1% Tween-20 or Triton X-100
Storage Solution: Deionized water, 0.05% Kv300
II. Bead Activation
Transfer 200 µL of UltraBio™ Carboxyl Magnetic Beads (1 μm) to a microcentrifuge tube. Vortex-mix thoroughly.
Place the tube on a magnetic rack for ~1 minute until the solution clears. Aspirate and discard the supernatant.
Note: Avoid aspirating beads. Applies throughout.
Remove the tube from the rack. Add an equal volume of Activation Solution. 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 Step 3 & 4 once more (1 wash total with Activation Solution).
Add 200 µL of freshly prepared EDC Solution (10 mg/mL). Mix thoroughly.
Incubate at room temperature for 30 minutes. Ensure the beads remain uniformly suspended to maximize activation efficiency.
Place the tube on the magnetic rack for ~1 minute. Aspirate and discard the supernatant.
Wash the beads twice rapidly with pre-chilled Deionized Water followed by two rapid washes with Activation Solution (Steps 3 & 4). Minimize time to prevent hydrolysis of activated groups.
III. Ligand Coupling
Add 200 µL of ligand solution (e.g., antibody/protein at 1-5 mg/mL in Activation Solution) to the activated beads. Mix thoroughly.
Incubate with end-over-end rotation at room temperature overnight (12-16 hours). Ensure the beads remain uniformly suspended.
Place the tube on the magnetic rack for ~1 minute. Aspirate and discard the supernatant.
Add 500 µL Blocking Solution. Incubate at room temperature for 30 minutes.
Aspirate the blocking solution. Wash beads twice with Wash Solution, followed by two washes with 1× PBS, pH 7.4.
Resuspend the coupled beads in Storage Solution for immediate use or store at 2-8°C for long-term storage.
IV. 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 |
|---|---|---|
 ZJ26F0737558 |
分析证书 | C1456040 |
| ZJ26F0737557 |
分析证书 | C1456040 |
| ZJ26F0131034 |
分析证书 | C1456040 |
| ZJ25F0826366 |
分析证书 | C1456040 |
| ZJ25F0826365 |
分析证书 | C1456040 |
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