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关于用EDC/NHS活化抗体上羧基的最佳条件的问题 已有4人参与
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各位虫友早上好，实验一直遇到瓶颈，感觉好烦躁，做电化学免疫传感器时为了将抗体结合在氨基化的无机材料上，用EDC/NHS活化的方法活化羧基，但是测DPV总感觉结合效果不好，不知道是不是活化条件没控制好，还是孵化过程有问题，希望做过相关工作并成效卓著的虫友能给个建议 发自小木虫IOS客户端 |
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2429650199
新虫 (正式写手)
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2楼2018-05-15 11:55:32
2429650199
新虫 (正式写手)
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3楼2018-05-15 11:55:41
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小木虫: 金币+0.5, 给个红包，谢谢回帖
小木虫: 金币+0.5, 给个红包，谢谢回帖
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4楼2018-05-18 19:40:21
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小木虫: 金币+0.5, 给个红包，谢谢回帖
小木虫: 金币+0.5, 给个红包，谢谢回帖
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Follow this paper to try periodate method to make orientated conjugation. Good luck. Sara Puertas, Pilar Batalla, María Moros, Ester Polo, Pablo del Pino, José M. Guisán, Valeria Grazú, Jesús M. de la Fuente. Taking Advantage of Unspecific Interactions to Produce Highly Active Magnetic Nanoparticle−Antibody Conjugates. ACS Nano. 2011, 5(6): 4521-4528. Just try the old classcial style (change its magnetic beads to your electrode surface) Immobilization via the Carbohydrates Moieties of the Antibody. One milliliter of an antibody solution 1 mg/mL in 10 mM sodium phosphate pH 7.0 was incubated with 100 μL of 0.1 M NaIO4 (prepared in water) during 2 h at 4C and preserved from light. Then, the oxidized antibody was purified by Sephadex G-25 gel filtration column with 10 mM sodium phosphate pH 8.0. Aliquots of 10 mg of aminated nanoparticles were incubated during 2 h at 37 C with 1 mL of a 100 μg/mL solution of oxidized antibody in 10 mM sodium phosphate pH 8.0. Then, NaCNBH3 was added in order to have a final concentration of 0.25 M. After 30 min at 37 C, the functionalized nanoparticles were washedthree times with10 mM sodium phosphate pH 8.0. Finally, the remaining amine groups of the nanoparticles were blocked by incubation with 1 mL BSA 1% in 10 mM MES pH 6.1 for 16 h at 25 C and were stored at 4 C until used. |
5楼2018-05-20 05:05:28
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小木虫: 金币+0.5, 给个红包，谢谢回帖
小木虫: 金币+0.5, 给个红包，谢谢回帖
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" Then, the oxidized antibody was purified by Sephadex G-25 gel filtration column with 10 mM sodium phosphate pH 8.0. " This step can be replaced using Zeba™ Spin Desalting Columns "https://www.thermofisher.com/ord ... ct/89882". Much simple and fast without diluting your antibody. |
6楼2018-05-20 05:16:30
7楼2018-05-22 10:23:46
8楼2018-06-29 16:05:39