琼脂糖基质磁性微球（Magnetic Agarose Beads）系列产品具有超顺磁性、快速磁响应性、丰富羟基官能团和相对集中的粒径等特点，能够在特殊化学试剂的作用下，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，是医学与分子生物学研究中重要的载体工具。本产品采用独特生产工艺制备的磁性琼脂糖微球，粒径分布在30-100 μm，粒径适中，更适合生物检测和纯化实验的需求。
链霉亲和素琼脂糖磁珠是将链霉亲和素（Streptavidin）通过化学方法固定在琼脂糖基质磁性微球表面，利用生物素与链霉亲和素之间的特异性相互作用，实现固定化生物素或生物素化蛋白、抗体等物质的目的。
| 项目 | 性能 |
|---|
| 基质 | 磁性琼脂糖微球 | | 配体 | 链霉亲和素 | | 结合能力 | ＞120 nmol Free biotin/ml 磁珠 | | 粒径范围 | 30-100 μm | | 储存缓冲液 | 含20%乙醇的1×PBS | | 磁珠体积 | 磁珠体积占悬浮液体积的20% | | 储存温度 | 2-8℃ |
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使用方法
1. 缓冲液准备
实验所用水和各类缓冲液，使用前建议用0.22 μm或0.45 μm滤膜过滤，去除杂质，避免影响实验效果。
(1) 生物素或生物素化物质的纯化
平衡/洗杂液：20 mM NaH₂PO₄，0.15 M NaCl，pH 7.4
洗脱液：8 M 盐酸胍，pH 1.5
(2) 亚氨基生物素标签物质的纯化
平衡/洗杂液：50 mM 碳酸铵，0.5 M NaCl，pH 10.0
洗脱液：50 mM 碳酸铵，0.5 M NaCl，pH 4.0
2. 样品准备
上柱前需确保样品溶液具有合适的离子强度和pH值，可采用以下两种方式调节：用对应平衡/洗杂液稀释血清样品、腹水或细胞培养液；或用对应平衡/洗杂液对样品进行透析。样品上样前，建议离心处理或用0.22μm滤膜过滤，去除样品中的杂质，提高蛋白纯化效率。
3. 操作流程
(1) 磁珠准备：将磁珠悬浮液反复颠倒，充分混匀；用移液器吸取适量磁珠悬浮液置于离心管中，将离心管放入磁分离器，静置约1 min，待溶液澄清后，用移液器吸弃上清液。
(2) 磁珠平衡：将离心管从磁分离器上取下，加入与磁珠悬浮液等体积的结合液，用枪头反复吹打5-10次，充分混匀；再次将离心管置于磁分离器，静置约1min，待溶液澄清后吸弃上清液；重复此洗涤步骤2次。
(3) 生物素化样品结合：以生物素化样品结合为例，将生物素化样品加入处理好的磁珠中，颠倒离心管充分混匀；将离心管置于混合仪上，室温孵育30 min以上（具体孵育时间可根据实际结合效果调整）。
(4) 洗杂：将离心管置于磁分离器，静置约1 min，待溶液澄清后，用移液器移出上清液并保留，以备后续取样检测；向离心管中加入5倍磁珠悬浮液体积的洗杂液，用枪头反复吹打5-10次，充分混匀；将离心管放回磁分离器，静置约1 min，待溶液澄清后吸弃上清液；重复此洗杂步骤2次以上。
(5) 洗脱：加入3-5倍磁珠体积的洗脱液，充分混合均匀，室温孵育5 min；将离心管置于磁分离器，待磁珠全部吸附后，吸取上清液（可重复洗脱2次，洗脱液可保留用于进一步检测）。
(6) 变性洗脱：将离心管从磁分离器上取下，向其中加入等体积2×SDS-PAGE Loading Buffer，充分混合均匀；95℃加热10 min，随后进行磁性分离；收集上清液，可通过SDS-PAGE检测或western blot方法检测目标分子。注意：此方法洗脱后，磁珠不可重复使用。
注意事项
1. 磁珠操作全程需轻柔，避免剧烈震荡，防止磁珠破损影响结合效果。
2. 孵育、磁分离时间可根据实验实际情况适当调整，确保磁珠充分吸附目标物质。
3. 变性洗脱后的磁珠因结构被破坏，禁止重复使用，避免影响后续实验结果。
4. 所有缓冲液需现配现用，或按要求储存，避免变质；储存温度需严格控制在2-8℃，防止磁珠失活。
Agarose Matrix Magnetic Microspheres (Magnetic Agarose Beads) series products feature superparamagnetism, fast magnetic response, abundant hydroxyl functional groups, and a relatively narrow particle size distribution. They can covalently couple biological ligands such as peptides, proteins, antibodies, and oligonucleotides to the microsphere surface under the action of specific chemical reagents, making them important carrier tools in medical and molecular biology research. This product is manufactured using a unique production process to produce magnetic agarose microspheres with a particle size distribution of 30–100 μm. The moderate particle size makes them more suitable for the needs of bioassays and purification experiments.
Streptavidin Agarose Magnetic Beads are prepared by chemically immobilizing streptavidin onto the surface of agarose matrix magnetic microspheres. They exploit the specific interaction between biotin and streptavidin to achieve immobilization of biotin or biotinylated proteins, antibodies, and other substances.
| Property | Description |
|---|
| Matrix | Magnetic agarose microspheres | | Ligand | Streptavidin | | Binding capacity | >120 nmol free biotin/mL beads | | Particle size range | 30–100 μm | | Storage buffer | 1× PBS containing 20% ethanol | | Bead volume | 20% of suspension volume | | Storage temperature | 2–8°C |
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Instructions for Use
1. Buffer preparation
All water and buffers used in the experiment should be filtered through a 0.22 μm or 0.45 μm filter before use to remove impurities that might affect the results.
(1) Purification of biotin or biotinylated substances
Equilibration/wash buffer: 20 mM NaH₂PO₄, 0.15 M NaCl, pH 7.4
Elution buffer: 8 M guanidine hydrochloride, pH 1.5
(2) Purification of iminobiotin‑labeled substances
Equilibration/wash buffer: 50 mM ammonium carbonate, 0.5 M NaCl, pH 10.0
Elution buffer: 50 mM ammonium carbonate, 0.5 M NaCl, pH 4.0
2. Sample preparation
Before loading, ensure the sample solution has appropriate ionic strength and pH. Adjust using one of the following methods: dilute serum samples, ascites, or cell culture media with the corresponding equilibration/wash buffer; or dialyze the sample against the corresponding equilibration/wash buffer. It is recommended to centrifuge the sample or filter it through a 0.22 μm filter before loading to remove impurities and improve protein purification efficiency.
3. Procedure
(1) Bead preparation: Invert the bead suspension repeatedly to mix thoroughly. Pipette an appropriate amount of bead suspension into a centrifuge tube. Place the tube in a magnetic separator and let stand for about 1 min until the solution becomes clear. Aspirate and discard the supernatant.
(2) Bead equilibration: Remove the tube from the magnetic separator. Add an equal volume of binding buffer (relative to the bead suspension volume) and pipette up and down 5–10 times to mix thoroughly. Place the tube back on the magnetic separator, let stand for about 1 min until the solution clears, then aspirate and discard the supernatant. Repeat this wash step twice.
(3) Binding of biotinylated sample (example): Add the biotinylated sample to the prepared beads. Invert the tube to mix thoroughly. Place the tube on a mixer and incubate at room temperature for at least 30 min (incubation time can be adjusted according to actual binding efficiency).
(4) Washing: Place the tube on the magnetic separator, let stand for about 1 min until the solution clears, then transfer the supernatant to a new tube (keep for subsequent analysis if needed). Add 5 volumes of wash buffer (relative to the bead suspension volume) to the tube and pipette up and down 5–10 times to mix thoroughly. Place the tube back on the magnetic separator, let stand for about 1 min until the solution clears, then aspirate and discard the supernatant. Repeat this washing step at least twice.
(5) Elution: Add 3–5 volumes of elution buffer (relative to the bead volume) and mix thoroughly. Incubate at room temperature for 5 min. Place the tube on the magnetic separator; after all beads are attracted, aspirate the supernatant (elution can be repeated twice, and the eluates can be retained for further analysis).
(6) Denaturing elution: Remove the tube from the magnetic separator. Add an equal volume of 2× SDS‑PAGE loading buffer and mix thoroughly. Heat at 95°C for 10 min, then perform magnetic separation. Collect the supernatant, which can be analyzed by SDS‑PAGE or western blot. Note: After denaturing elution, the beads cannot be reused.
Precautions
Always handle the beads gently throughout the procedure; avoid vigorous shaking to prevent bead damage that could affect binding efficiency.
Incubation and magnetic separation times may be adjusted according to the actual experimental conditions to ensure sufficient capture of target substances.
Beads subjected to denaturing elution are structurally damaged and must not be reused, as this may compromise subsequent experimental results.
All buffers should be freshly prepared or stored as required to avoid degradation. Storage temperature must be strictly controlled at 2–8°C to prevent bead inactivation.
ZJ26F0737827
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