本文選題：SMO + 重組蛋白�。� 參考：《三峽大學(xué)》2012年碩士論文

【摘要】：目的：制備鼠抗人的精胺氧化酶(Spermine oxidase, SMO)單克隆抗體，分析該抗體的效價、抗原特異性、種屬特異性及在Western blot，免疫組織化學(xué)，免疫沉淀，免疫熒光，流式細(xì)胞術(shù)等分析技術(shù)中的可用性。 方法：在BL21(DE3)中原核表達(dá)帶6×His標(biāo)簽的SMO重組蛋白，并用Ni-NTA樹脂親和層析純化。用SMO重組蛋白免疫Balb/C小鼠，通過雜交瘤技術(shù)制備抗SMO單克隆抗體，所得抗體的特異性及效價用ELISA和Western blot以及免疫組織化學(xué)等技術(shù)檢測。 結(jié)果：成功建立了一株穩(wěn)定分泌抗人SMO的單克隆抗體雜交瘤細(xì)胞株，該單克隆抗體可用于ELISA，Western blot，免疫組織化學(xué)，免疫熒光，免疫沉淀，，流式細(xì)胞術(shù)等分析技術(shù)。 結(jié)論：成功制備了高效價的SMO單克隆抗體，為SMO相關(guān)的分子、病理及抗腫瘤研究奠定了基礎(chǔ)。
[Abstract]:Objective: to prepare mouse monoclonal antibody against human spermine oxidase (SMOA), and analyze its titer, antigen specificity, species specificity, immunocytochemistry, immunoprecipitation and immunofluorescence. Availability in analytical techniques such as flow cytometry. Methods: SMO recombinant protein with 6 脳 His tag was expressed in prokaryotic cells of BL21 DDE3 and purified by Ni-NTA resin affinity chromatography. SMO recombinant protein was used to immunize Balb/C mice. Monoclonal antibodies against SMO were prepared by hybridoma technique. The specificity and titer of the antibodies were detected by ELISA, Western blot and immunohistochemistry. Results: a hybridoma cell line with stable secretion of anti-human SMO was successfully established. The monoclonal antibody can be used for the analysis of ELISAL Western blot, immunohistochemistry, immunofluorescence, immunoprecipitation, flow cytometry and so on. Conclusion: the SMO monoclonal antibody with high titer was successfully prepared, which laid a foundation for the study of SMO related molecules, pathology and antitumor.
【學(xué)位授予單位】：三峽大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R392.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 王艷林;韓鈺;袁太寧;曹春雨;周永琴;;多胺類似物CPENSpm通過干擾多胺代謝抑制肺癌細(xì)胞的增殖[J];中國藥理學(xué)通報;2008年01期

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