计算溶液所需的质量、体积或浓度。
为什么选择此级别
，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
2-8°C储存,避光。低温运输 。请查阅批次 COA 获取详细规格。
质量文档
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。
文献证明
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。
概述
本产品是含有稀土元素 EU 的聚苯乙烯微球。激发波长 300~500nm(常用 365nm），发射光具有 stokes 位移大、荧光寿命长、检测信号强等特点。与常规荧光相比，本产品荧光信号强度高，更适用于层析定量高灵敏项目的开发以及生产。
使用方法
一、一步法标记工艺
1、取出微球原液超声分散。
2、抗体的偶联过程。
以标记 200ul 体积为例：取 20ul (10mg/ml)时间分辨荧光微球加入到 180ul0.01MPB pH6.2 的缓冲液中，震荡混匀后，加入终浓度 150ug/ml 的抗体，随后加入 10ulEDC(0.1 mg/ml)，混匀震荡反应2h。（注：EDC 用 0.05M MES pH5.5 的缓冲液溶解；标记缓冲液应适当调整，说明书涉及的缓冲液仅供参考）
3、封闭：加入适量封闭液，室温震荡反应 0.5~1h。
加 100ul 5%BSA 水溶液，室温垂直混匀(封闭)0.5~1h。
4、重悬：离心，去除上清后，加入适量复溶液复溶至定量浓度，即可使用。
10000prm 离心 10min,去除上清后，加入 500ul 复溶液定容到 0.4mg/ml，超声分散，4℃保存；
二、两步法标记工艺
1、取出微球原液超声分散。
2、微球活化: 加入 EDC 、NHS 活化液反应 0.5~1h。
以标记 200ul 体积为例：取 20ul (10mg/ml)时间分辨荧光微球加入到 180ul 含有 0.05%TW20 的0.05M MES pH5.5 缓冲液中，震荡混匀后，加入 2ulNHS(75 mg/ml)、2ulEDC(50 mg/ml)，混匀震荡反应0.5h。（注：EDC、NHS 均用 0.05M MES pH5.5 的缓冲液溶解）
3、重悬：离心，去上清，后用合适的缓冲液复溶，超声分散。
离心条件：10000rpm 离心 10min，去上清后加 200ul 0.05M MES5.5 重悬，超声分散。（离心条件应随反应体积的变化适当调整，标记缓冲液应根据需求调整）
4、偶联：向以上加入适量的待标记抗体、抗原、核酸等，震荡反应 1~2h。在反应过程中若出现聚集情况，需超声分散。
加入 150ug/ml 抗体，偶联 1h。（若需要独立标记，则 DNP-BSA 标记浓度 150ug/ml，再反应 1h）
5、封闭：加入适量封闭液，室温震荡反应 0.5~1h。
加 100ul 5%BSA 的 Tris 溶液(0.1M Ph8.0)，室温垂直混匀(封闭)0.5h:
6、重悬：离心，去除上清后，加入适量复溶液复溶至定量浓度，即可使用。
10000prm 离心 10min,去除上清后，加入 500ul 复溶液定容到 0.4mg/ml，超声分散，4℃保存；
注意事项
1、缓冲液使用不合适，会出现标记聚集的情况，建议在标记过程中增加超声操作，分散微球。
2、离心条件应随离心体积的变化做出相应的调整。
3、荧光微球使用之前，需超声分散。
4、偶联效率受 pH、温度、分子量、盐浓度的影响，建议优化反应条件。
5、废弃物应按照危险品处理。
产品优势
1. 表面功能化：羧基；
2. 荧光稳定性更佳；
3. 荧光微球粒径均一，单分散性优异，稳定性好，PDI系数均≤0.3；
4. 检测灵敏度更高；
5. 适配免疫层析定量检测、微孔板超敏检测两大技术平台。
This product is polystyrene microspheres doped with rare earth element europium (Eu). It has an excitation wavelength of 300–500 nm (365 nm is commonly used), and its emitted fluorescence features a large Stokes shift, long fluorescence lifetime and strong detection signals. Compared with conventional fluorescent materials, this product delivers high fluorescence signal intensity, making it more suitable for the development and mass production of high-sensitivity quantitative lateral flow assays.
Usage
I. One-Step Labeling Process
1. Remove the microsphere stock solution and disperse it using ultrasound.
2. Antibody coupling process.
Using a 200 μl volume for labeling as an example: Add 20 μl of (10 mg/ml) time-resolved fluorescent microspheres to 180 μl of 0.01 M PB (pH 6.2). After shaking to mix, add the antibody to a final concentration of 150 μg/ml, followed by 10 μl of EDC (0.1 mg/ml). Mix well and incubate with shaking for 2 hours. (Note: Dissolve EDC in 0.05 M MES buffer (pH 5.5); the labeling buffer should be adjusted appropriately; the buffers mentioned in the manual are for reference only)
3. Blocking: Add an appropriate amount of blocking solution and incubate with shaking at room temperature for 0.5–1 hour.
Add 100 μl of a 5% BSA aqueous solution and mix vertically at room temperature (blocking) for 0.5–1 hour.
4. Resuspension: Centrifuge, remove the supernatant, then add an appropriate amount of resuspension buffer to resuspend to the desired concentration; the sample is now ready for use.
Centrifuge at 10,000 rpm for 10 minutes, remove the supernatant, add 500 μl of resuspension buffer to adjust the concentration to 0.4 mg/ml, disperse by sonication, and store at 4°C;
II. Two-Step Labeling Procedure
1、Take out the microsphere stock solution and subject it to ultrasonic dispersion.
2、 Microsphere activation: Add EDC and NHS activation solution and react for 0.5~1h.
Taking the labeling of a 200ul volume as an example: Take 20ul (10mg/ml) of time-resolved fluorescent microspheres and add them to 180ul of 0.05M MES pH5.5 buffer containing 0.05% TW20. After shaking and mixing, add 2ul NHS (75 mg/ml) and 2ul EDC (50 mg/ml), and mix and shake for a reaction time of 0.5h. (Note: Both EDC and NHS are dissolved in 0.05M MES pH5.5 buffer.)
3、Resuspension: Centrifuge, remove the supernatant, and then resuspend with an appropriate buffer solution, followed by ultrasonic dispersion.
Centrifugation conditions: centrifuge at 10,000 rpm for 10 minutes, remove the supernatant, add 200 μl of 0.05M MES5.5 for resuspension, and then perform ultrasonic dispersion. (Centrifugation conditions should be adjusted appropriately according to changes in reaction volume, and the labeling buffer should be adjusted as needed.)
4、Conjugation: Add an appropriate amount of antibodies, antigens, nucleic acids, etc. to the above solution, and shake the reaction for 1-2 hours. If aggregation occurs during the reaction, perform ultrasonic dispersion.
Add 150 μg/ml antibody and conjugate for 1 hour. (If independent labeling is required, use a DNP-BSA labeling concentration of 150 μg/ml and react for another 1 hour.)
5、Blocking: Add an appropriate amount of blocking solution and shake at room temperature for 0.5~1h.
Add 100ul of 5% BSA in Tris solution (0.1M, pH 8.0) and mix vertically at room temperature for 0.5h (blocking).
6、Resuspending: Centrifuge, remove the supernatant, and add an appropriate amount of resuspension solution to resuspend to a quantitative concentration for use.
Centrifuge at 10000prm for 10min, remove the supernatant, add 500ul of resuspension solution to dilute to 0.4mg/ml, ultrasonicate for dispersion, and store at 4℃;
Precautions
1.Inappropriate use of buffer solution may lead to aggregation of markers. It is recommended to add ultrasonic operation during the marking process to disperse the microspheres.
2.Centrifugation conditions should be adjusted accordingly with changes in centrifuge volume.
3.Fluorescent microspheres need to be ultrasonically dispersed before use.
4.Coupling efficiency is affected by pH, temperature, molecular weight, and salt concentration. It is recommended to optimize the reaction conditions.
5. Waste should be disposed of as hazardous waste.
Product Advantage
1.Surface functionalization: carboxyl groups；
2.Better fluorescence stability；
3.Fluorescent microspheres have uniform particle size, excellent monodispersity and good stability,The PDI index is all ≤ 0.3；
4.Higher detection sensitivity；
5.Suitable for two major technical platforms: quantitative immunochromatographic detection and microplate ultrasensitive detection.
规格
规格或纯度
粒径：300nm；表面官能团：-COOH；10mg/ml in H2O
英文名称
Time-resolved fluorescent microspheres
储存条件
2-8°C储存,避光
运输条件
低温运输
名称和识别符
分子类型
未知
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):
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技术文档和文章
溶液计算器
产品仅提供给经过验证的企业、机构和合格的专业人员用于研发。不适用于人类、动物、诊断或治疗。
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