| R-藻红蛋白免疫荧光探针标记方法的探索 |
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| 引用本文： | 王盛,钟伏弟,吴祖建,林奇英,谢联辉.R-藻红蛋白免疫荧光探针标记方法的探索[J].福建农林大学学报(自然科学版),2004,33(2):206-209. |
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| 作者姓名： | 王盛 钟伏弟 吴祖建 林奇英 谢联辉 |
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| 作者单位： | 福建农林大学植物病毒研究所,福建,福州,350002 |
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| 基金项目： | 福建省科技厅重大资助项目(2001Z127). |
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| 摘 要： | 利用双功能试剂SPDP在蛋白质分子中引入巯基,结合2-巯醇吡啶分析方法,定量测定引入活性基团的比例,通过两步标记法,将珊瑚藻R-藻红蛋白标记到羊抗兔抗体上,简化了传统标记过程.参照Dot-ELISA方法,以硝酸纤维素膜为固相载体,将该方法应用于烟草花叶病毒(Tobaccomosaicvirus,TMV)的检测.结果表明,R-PE标记的荧光检测的灵敏度为2-10,与酶免疫的Dot-ELISA相比,尽管检测灵敏度较低,但快捷、经济.并对如何进一步提高藻红蛋白介导的免疫荧光分析方法的灵敏度作了讨论.
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| 关 键 词： | 藻红蛋白 荧光探针 标记 |
| 文章编号： | 1006-7817(2004)02-0206-04 |
| 修稿时间： | 2003年10月20 |
Research on R-phycoerythrin fluorescently-labeled probes |
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| WANG Sheng,ZHONG Fu-di,WU Zu-jian,LIN Qi-ying,XIE Lian-hui.Research on R-phycoerythrin fluorescently-labeled probes[J].Journal of Fujian Agricultural and Forestry University,2004,33(2):206-209. |
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| Authors: | WANG Sheng ZHONG Fu-di WU Zu-jian LIN Qi-ying XIE Lian-hui |
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| Abstract: | By means of heterobifunctional crosslinker-SPDP, sulfhydryl could be added to a protein, and the ratio also could be calculated using protocol for Pyridine-2-Thione assay. Based on this, we prepared the phycoerythrin-sheep anti-rabbit IgG flour probes by two-step binding protocol, which protocol was easier than tradition′s. Using nylon membranes as solid-phase medium, we applied the protocol of dot-ELISA to detecting Tobacco mosaic virus(TMV). The result showed that the sensitivity of immunoassay was 2-10 by using R-PE as probes, and was lower than that in assay of dot-ELISA, but it was more convenient and economical. We also discussed how to improve the sensitivity of immunoassay by using the phycoerythrin flour as probes. |
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| Keywords: | phycoerythrin flour probe label |
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