计算溶液所需的质量、体积或浓度。
羧基磁珠
只有 %1 左
概述
磁微粒化学发光免疫分析是将磁性分离技术、化学发光技术、免疫分析技术三者结合的分析方法。磁微粒作为载体偶联抗体或抗原，可捕获目标分子，因其具有超顺磁性可实现多次磁分离，有利于检测仪器的全自动化并加快检测速度。目前，磁微粒化学免疫分析技术因其灵敏度高、检测范围宽、自动化程度高等优点已被广泛应用于生物医学检测领域。
羧基磁珠表面有羧基（COOH）修饰，在碳化二亚胺（EDC）活化下可与蛋白共价结合。EDC反应形成的中间体在水中不稳定，容易水解，为了防止活化后的中间体快速水解，提高偶联效率，可在反应中添加N- 羟基琥珀酰亚胺磺酸钠（Sulfo-NHS），以形成更稳定的中间体。
| 参数 | C1506716-1.2μm | C1506716-2.8μm |
| 外观 | 红棕色悬浊液 | 红棕色悬浊液 |
| 固含量 | 10 mg/mL | 10 mg/mL |
| 粒径及分布 | 1.2± 0.1 μm, CV＜5% | 2.8± 0.1 μm, CV＜5% |
| 表面基团含量 | 200-300 μmol/g | 200-300 μmol/g |
产品特点
1、弥散式结构：磁核材料为超顺磁性γ-Fe₂O₃纳米颗粒
2、氧化铁含量：12%-20%（不同磁珠有差异）
3、粒径：常规1.2、2.8μm
4、 粒径高度均一，批间差CV＜5%：保证稳定可靠的性能
5、分散性优良，检测本底低：自主创新设计的独特包覆层结构，具有极低的非特异性吸附
6、单批次2公斤级产能：满足化学发光客户日常需求
7、高稳定性：长时间存储后不会出现氧化变色和核磁泄露现象
使用说明
1、一步法偶联与两步法偶联选择
一步偶联法相较于两步法更简单易操作，但由于偶联蛋白 （抗体、抗原等）本身含有伯胺及羧基基团，其在与EDC反应时易产生自身交联。
两步偶联法有Sulfo-NHS参与反应的过程中会形成更稳定的中间体 , 活化后的清洗过程可去除过量的 EDC，可以消除偶联蛋白自身的羧基基团被活化而产生蛋白自身交联的可能性。
两步偶联法有助于在偶联过程中更换偶联缓冲液，通过筛选不同的偶联缓冲液及条件，以提高蛋白偶联效率。
建议初始偶联方案可以尝试一步法和两步法，以筛选最佳偶联方案。
2、活化条件选择
(1) 活化过程中使用的缓冲液不应含有伯胺或是羧基基团，例如 Tris 缓冲液、甘氨酸缓冲液、柠檬酸缓冲液等。
(2) EDC与Sulfo-NHS在偏酸性条件下活化效率高，推荐使用 MES（2- 吗啉乙磺酸）缓冲液，pH 4.5~6.5。
(3) 在活化磁珠时EDC与Sulfo-NHS应使用活化缓冲液（例 MES 缓冲液）现配现用。
3、偶联条件选择
(1) 偶联缓冲液不应含有伯胺基团，例如 Tris 缓冲液、甘氨酸缓冲液等。
(2) 一步偶联法通常活化过程与偶联过程在同缓冲液中进行，同时需考虑 COOH 最优活化条件以及蛋白偶联最优条件，推荐使用 MES 缓冲液，pH 4.5~6.5。
(3) 两步偶联法推荐MES（pH 5.0）或PBS（pH 7.2）作为偶联缓冲液，建议可尝试 pH 4.5~7.5，以筛选最佳偶联条件。
(4) 以IgG抗体为例，推荐偶联比例为IgG:Beads=20~30μg:1mg；但由于偶联蛋白的多样性，建议筛选不同的偶联比例，以获得最优结果。
(5) 一步法与两步法推荐偶联时长为2~6h，建议尝试不同偶联时长，以筛选最佳偶联条件。
(6) 偶联完成后建议使用BSA、酪蛋白等蛋白或其他合成封闭剂进行封闭。
4、偶联注意事项
(1) EDC 易水解，受潮或结块后不建议使用，需要保存在 -20℃干燥容器内，使用前平衡至室温。
(2) 磁珠在取用前，务必充分振荡使磁珠呈均匀的悬浮状态。
(3) 干燥、冷冻操作极易引起磁珠团聚，且会影响磁珠表面功能基团的活性，注意保存条件。
5、偶联推荐方案
| 名称 | 详述 | 配制方法简述 |
| 偶联缓冲液 | 25mM MES缓冲液, pH 5.0 | 0.533g MES (2-吗啉乙磺酸单水合物, MW 213.25)，使用80mL纯化水溶解，调节pH到5.0，补加纯化水至100mL |
| 封闭液 | 50mM Tris+1.0% BSA+0.1% Tween 20, pH 7.4 | 0.606g Tris (三羟甲基氨基甲烷, MW 121.14)，1.0g BSA，0.1g Tween20使用80mL纯化水溶解，调节pH到7.4，补加纯化水至100mL |
| 保存液 | 50mM Tris+1.0% BSA+0.1% Tween 20, pH 7.4 | 同上 |
6、一步法
(1) 使用涡旋混匀器将羧基磁珠充分混匀悬浮，取10mg到离心管中；
(2) 将离心管放置在磁力架上1~2min，移除上清液；
(3) 加入1.0mL的偶联缓冲液，涡旋分散磁珠后，按照步骤 (2) 移除上清；
(4) 重复 (3) 步骤2次；
(5) 加入200μg抗体，补加偶联缓冲液至总体积为0.9mL，涡旋分散磁珠；
(6) 保持磁珠混悬状态，在室温条件下反应15~30min；
(7) 加入EDC溶液0.1mL（10mg/mL，用偶联缓冲液现配现用），保持磁珠混悬状态，在室温条件下反应2~6h后按照步骤 (2) 移除上清；
(8) 加入1.0mL封闭液，涡旋分散磁珠；
(9) 保持磁珠混悬状态，在室温或 37℃条件下2~6h，按照步骤 (2) 移除上清；
(10) 加入1.0mL保存液，涡旋分散磁珠后，按照步骤 (2) 移除上清；
(11) 重复 (10) 步骤2次；
(12) 加入1.0mL保存液，涡旋混匀，得到浓度为10mg/mL的磁珠混悬液，2~8℃下保存。
7、两步法
(1) 使用涡旋混匀器将羧基磁珠充分混匀悬浮，取10mg到离心管中；
(2) 将离心管放置在磁力架上1~2min，移除上清液；
(3) 加入1.0mL的偶联缓冲液，涡旋分散磁珠后，按照步骤 (2) 移除上清；
(4) 重复 (3) 步骤2次；
(5) 加入EDC溶液0.1mL（10mg/mL，用偶联缓冲液现配现用）和 Sulfo-NHS 溶液0.1mL（10mg/mL，用偶联缓冲液现配现用），补加偶联缓冲液至总体积为1.0mL，涡旋分散磁珠；
(6) 保持磁珠混悬状态，在室温条件活化反应30min后按照步骤 (2) 移除上清液；
(7) 加入200μg抗体，补加偶联缓冲液至总体积为1.0mL，涡旋分散磁珠；
(8) 保持磁珠混悬状态，在室温条件下反应2~6h后按照步骤 (2) 移除上清；
(9) 加入1.0mL封闭液，涡旋分散磁珠；
(10) 保持磁珠混悬状态，在室温或37℃条件下2~6h，按照步骤 (2) 移除上清；
(11) 加入1.0mL保存液，涡旋分散磁珠后，按照步骤 (2) 移除上清；
(12) 重复步骤 (11) 2次；
(13) 加入1.0mL保存液，涡旋混匀，得到浓度为10mg/mL的磁珠混悬液，2~8℃下保存。
应用实例
1、C1506716-1.2μm在TSH检测中与进口品牌D发光值对比
结论：
相同的磁珠偶联、试剂检测条件下，
C1506716-1.2μm磁珠在TSH检测中发光值与进口品牌D同类型1.0μm磁性微球基本一致；
C1506716-1.2μm三批发光值差异极小，生产工艺稳定批间差小。
2、C1506716-2.8μm在AFP检测中与进口品牌D发光值对比
结论：
相同的磁珠偶联、试剂检测条件下，
C1506716-2.8μm磁珠在AFP检测中发光值高于进口品牌D同类型2.8μm磁性微球；
C1506716-2.8μm三批发光值差异极小，生产工艺稳定批间差小。
Magnetic Particle Chemiluminescence Immunoassay is an analytical method that combines magnetic separation technology, chemiluminescence technology, and immunoassay technology. Magnetic particles, as carriers conjugated with antibodies or antigens, can capture target molecules. Due to their superparamagnetic properties, they enable multiple magnetic separations, facilitating full automation of detection instruments and accelerating testing speed. Currently, magnetic particle chemiluminescence immunoassay technology is widely used in biomedical detection due to its high sensitivity, broad detection range, and high automation.
Carboxyl Magnetic Beads are surface-modified with carboxyl groups (COOH), which can covalently bind to proteins under activation by carbodiimide (EDC). The intermediate formed by the EDC reaction is unstable in water and prone to hydrolysis. To prevent rapid hydrolysis of the activated intermediate and improve conjugation efficiency, sodium N-hydroxysulfosuccinimide (Sulfo-NHS) can be added to the reaction to form a more stable intermediate.
| Parameter | C1506716-1.2μm | C1506716-2.8μm |
| Appearance | Red-brown suspension | Red-brown suspension |
| Solid Content | 10 mg/mL | 10 mg/mL |
| Particle Size & Distribution | 1.2± 0.1 μm, CV＜5% | 2.8± 0.1 μm, CV＜5% |
| Surface Group Content | 200-300 μmol/g | 200-300 μmol/g |
Product Features
Diffuse Structure: The magnetic core consists of superparamagnetic γ-Fe₂O₃ nanoparticles.
Iron Oxide Content: 12% - 20% (varies depending on the specific bead type).
Particle Size: Standard sizes of 1.2 μm and 2.8 μm.
Highly Uniform Particle Size with CV < 5% between Batches: Ensures stable and reliable performance.
Excellent Dispersion and Low Detection Background: Features a uniquely designed, proprietary coating layer that exhibits extremely low non-specific adsorption.
Production Capacity of 2 kg per Batch: Meets the routine demands of chemiluminescence customers.
High Stability: No oxidation/discoloration or magnetic core leakage occurs after long-term storage.
Instructions for Use
1. One-Step vs. Two-Step Conjugation
The one-step method is simpler and more straightforward than the two-step method. However, since the conjugated protein (antibody, antigen, etc.) itself contains primary amine and carboxyl groups, it may lead to self-crosslinking when reacting with EDC.
In the two-step method, the involvement of Sulfo-NHS forms a more stable intermediate. The washing step after activation removes excess EDC, eliminating the possibility of self-crosslinking due to activation of the protein's own carboxyl groups.
The two-step method allows for changing the conjugation buffer during the process, enabling optimization of conjugation efficiency by screening different buffers and conditions.
It is recommended to try both one-step and two-step methods initially to determine the optimal conjugation protocol.
2. Activation Conditions
(1) The buffer used during activation should not contain primary amines or carboxyl groups, such as Tris buffer, glycine buffer, or citrate buffer.
(2) EDC and Sulfo-NHS exhibit high activation efficiency under slightly acidic conditions. MES (2-morpholinoethanesulfonic acid) buffer at pH 4.5–6.5 is recommended.
(3) When activating magnetic beads, EDC and Sulfo-NHS should be prepared fresh using the activation buffer (e.g., MES buffer).
3. Conjugation Conditions
(1) The conjugation buffer should not contain primary amine groups, such as Tris buffer or glycine buffer.
(2) For the one-step method, activation and conjugation are typically performed in the same buffer. Consider the optimal activation conditions for COOH and the optimal conjugation conditions for proteins. MES buffer at pH 4.5–6.5 is recommended.
(3) For the two-step method, MES (pH 5.0) or PBS (pH 7.2) is recommended as the conjugation buffer. It is advisable to test pH 4.5–7.5 to determine the optimal conjugation conditions.
(4) For IgG antibodies, the recommended conjugation ratio is IgG:Beads = 20–30 μg:1 mg. However, due to the diversity of conjugated proteins, it is recommended to test different ratios for optimal results.
(5) For both one-step and two-step methods, the recommended conjugation duration is 2–6 hours. Testing different durations is advised to determine the optimal conditions.
(6) After conjugation, blocking with proteins such as BSA or casein, or other synthetic blocking agents, is recommended.
4. Conjugation Precautions
(1) EDC is prone to hydrolysis. Do not use if dampened or clumped. Store at -20°C in a dry container and equilibrate to room temperature before use.
(2) Before use, thoroughly vortex the magnetic microspheres to ensure a uniform suspension.
(3) Drying or freezing can cause aggregation of magnetic microspheres and affect the activity of surface functional groups. Pay attention to storage conditions.
5. Recommended Conjugation Protocol
| Item | Details | Preparation Method |
| Conjugation Buffer | 25 mM MES buffer, pH 5.0 | Dissolve 0.533 g MES (MW 213.25) in 80 mL purified water, adjust pH to 5.0, and add purified water to 100 mL. |
| Blocking Buffer | 50 mM Tris + 1.0% BSA + 0.1% Tween 20, pH 7.4 | Dissolve 0.606 g Tris (MW 121.14), 1.0 g BSA, and 0.1 g Tween 20 in 80 mL purified water, adjust pH to 7.4, and add purified water to 100 mL. |
| Storage Buffer | 50 mM Tris + 1.0% BSA + 0.1% Tween 20, pH 7.4 | Same as above. |
6. One-Step Method
(1) Thoroughly vortex the carboxyl magnetic microspheres to suspend evenly, and transfer 10 mg to a centrifuge tube.
(2) Place the tube on a magnetic rack for 1–2 minutes and remove the supernatant.
(3) Add 1.0 mL of conjugation buffer, vortex to disperse the microspheres, and remove the supernatant as in step (2).
(4) Repeat step (3) twice.
(5) Add 200 μg of antibody and supplement with conjugation buffer to a total volume of 0.9 mL. Vortex to disperse the microspheres.
(6) Maintain the microspheres in suspension and react at room temperature for 15–30 minutes.
(7) Add 0.1 mL of EDC solution (10 mg/mL, prepared fresh in conjugation buffer). Maintain suspension and react at room temperature for 2–6 hours. Remove the supernatant as in step (2).
(8) Add 1.0 mL of blocking buffer and vortex to disperse the microspheres.
(9) Maintain suspension and react at room temperature or 37°C for 2–6 hours. Remove the supernatant as in step (2).
(10) Add 1.0 mL of storage buffer, vortex to disperse, and remove the supernatant as in step (2).
(11) Repeat step (10) twice.
(12) Add 1.0 mL of storage buffer, vortex to mix, and store the magnetic microsphere suspension (10 mg/mL) at 2–8°C.
7. Two-Step Method
(1) Thoroughly vortex the carboxyl magnetic microspheres to suspend evenly, and transfer 10 mg to a centrifuge tube.
(2) Place the tube on a magnetic rack for 1–2 minutes and remove the supernatant.
(3) Add 1.0 mL of conjugation buffer, vortex to disperse the microspheres, and remove the supernatant as in step (2).
(4) Repeat step (3) twice.
(5) Add 0.1 mL of EDC solution (10 mg/mL, prepared fresh in conjugation buffer) and 0.1 mL of Sulfo-NHS solution (10 mg/mL, prepared fresh in conjugation buffer). Supplement with conjugation buffer to a total volume of 1.0 mL. Vortex to disperse the microspheres.
(6) Maintain suspension and activate at room temperature for 30 minutes. Remove the supernatant as in step (2).
(7) Add 200 μg of antibody and supplement with conjugation buffer to a total volume of 1.0 mL. Vortex to disperse the microspheres.
(8) Maintain suspension and react at room temperature for 2–6 hours. Remove the supernatant as in step (2).
(9) Add 1.0 mL of blocking buffer and vortex to disperse the microspheres.
(10) Maintain suspension and react at room temperature or 37°C for 2–6 hours. Remove the supernatant as in step (2).
(11) Add 1.0 mL of storage buffer, vortex to disperse, and remove the supernatant as in step (2).
(12) Repeat step (11) twice.
(13) Add 1.0 mL of storage buffer, vortex to mix, and store the magnetic microsphere suspension (10 mg/mL) at 2–8°C.
Application Examples
1. Comparison of Luminescence Values between C1506716-1.2μm and Imported Brand D in TSH Detection
Conclusion:
Under identical bead conjugation and reagent detection conditions:
The luminescence values of C1506716-1.2μm magnetic beads in TSH detection are essentially consistent with those of the similar 1.0μm magnetic microspheres from Imported Brand D.
The luminescence values across three batches of C1506716-1.2μm show minimal variation, indicating a stable production process and low batch-to-batch variation.
2. Comparison of Luminescence Values between C1506716-2.8μm and Imported Brand D in AFP Detection
Conclusion:
Under identical bead conjugation and reagent detection conditions:
The luminescence values of C1506716-2.8μm magnetic beads in AFP detection are higher than those of the similar 2.8μm magnetic microspheres from Imported Brand D.
The luminescence values across three batches of C1506716-2.8μm show minimal variation, indicating a stable production process and low batch-to-batch variation.
规格
规格或纯度
BioReagent, 用于化学发光, 10mg/mL
稳定性与储存
Store at 2-8℃ long term (48 months). Do not freeze. Keep the vial upright.
英文名称
Carboxyl Magnetic Beads
储存条件
2-8°C储存,禁止冷冻
运输条件
低温运输,禁止冷冻
名称和识别符
分子类型
生物试剂/缓冲液
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
通过匹配包装上的批号来查找并下载产品的 COA，每批产品都进行了严格的验证，您可放心使用！
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0737832 |
分析证书 | C1506716 |
| ZJ26F0737831 |
分析证书 | C1506716 |
| ZJ26F0535120 |
分析证书 | C1506716 |
| ZJ26F0535119 |
分析证书 | C1506716 |
| ZJ26F0535118 |
分析证书 | C1506716 |
| ZJ26F0535117 |
分析证书 | C1506716 |
技术文档和文章
溶液计算器
常见问题
本产品应如何储存？
请于2–8 °C条件下保存。请勿冷冻。冷冻可能对本品造成不可逆的损坏。
什么是「用于化学发光」？
「用于化学发光」表示该化学品经过纯化和检测，专用于专项分析技术。关键规格通常包括：常用检测波长处的受控紫外吸收、低不挥发残渣、受控的水分含量以及经验证的批间一致性。「用于化学发光」用于此工作流程中的流动相、样品制备和分析标准品。
什么是「BioReagent」？
「BioReagent」表示该产品经过加工和检测，适用于生物和生化应用。规格可能包括内毒素限值、微生物控制、无菌性、低 DNase/RNase/蛋白酶活性，以及在相关情况下的生物活性验证。
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