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摘要：本文制备了胶体Au和RE微粒（铈、钐、镧、钆、铕、钕、镝、铒）,研究了硝酸银浓度和RE微粒对银染效果影响。用胶体Au制备了TNNI3K浓度的检测试剂盒，研究了胶体Au标记一抗的浓度、胶体Au标记二抗的条件、 加入Au-二抗后的清洗次数、不同RE微粒与胶体Au混合比例对TNNI3K检测中银染效果影响。分别采用UV-Vis分光光度法和IR法对胶体Au、Au-二抗、RE微粒、RE-Au微粒、Au-二抗和TNNI3K浓度的检测试剂盒进行了表征。33668
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研究结果表明，硝酸银浓度为3.0×10-2mol/L、对二苯酚0.15mol/L时，银染效果好;与胶体Au相比，RE-Au微粒可延长银染后的斑点持续的时间，其中Nd-Au微粒的效果最好，斑点持续的时间为30mins，是胶体Au的2.7倍，可大幅度加深斑点的颜色，提高了银染法的灵敏度。当胶体金溶液的pH为6，1mL胶体金加入蛋白量为6.6µg的羊抗鼠TNNI3K二抗时，胶体Au与二抗结合形成的Au-二抗稳定；最佳一抗的浓度为10µg/mL，镧、钐、钕与胶体Au的混合比例（体积比）分别为7:3、5:5、6:4时，灰度值高。本文创新之处：RE-Au微粒应用在TNNI3K浓度的检测中未见报道。
毕业论文关键词：胶体Au；RE微粒；银染；
Application of gold particles in TNNI3K detection
Abstract: In this paper, colloidal Au and RE particles（La、Ce、Nd、Sm、Eu、Gd、Dy ）were prepared ，and their effects on silver staining were studied，a kits for determining TNNI3K concentration has been prepared with colloidal Au, The condition of labeling secondary antibody and washing time after join secondary antibody and the influence of mouse anti-human TNNI3K monoclonal antibody (primary antibody) concen- tration and the mixing proportion of RE-Au on the grey value were studied.UV-Vis and IR spectro were used to characterize colloidal Au, RE particles, RE-Au particles, Au-secondary antibody (Goat anti-mouse IgG(H+L) antibody: secondary antibody), and RE-Au-secondary antibody (RE: La, Sm, Nd) respectively.
The results showed that Silver nitrate concentration is 3.0×10-2mol/L, for phenol is 0.15mol/L silver dyeing effect is good Compared with colloidal Au, RE-Au particles could prolong the duration of the spots after silver staining. And Nd-Au showed the best effect that the duration time of the spots was 30mins which was 2.7 times longer than that of colloidal Au. It could greatly darken the color of spots. In addition, it could improve the sensitivity of silver staining by replacing a portion of colloidal Au. Au-secondary antibody was stable when adding 6.6μg secondary antibody to colloidal Au (1mL) of pH 6 for the kit (Au) for determining TNNI3K concentration; When the mixing proportion of RE:Au were 7:3, 5:5 and 6:4 respectively, the grey value was higher for the kits (RE-Au) for determining TNNI3K concentration; Innovation points of the research: the preparation of kits for determining TNNI3K concentration with RE-Au particles by filtration assay have not been reported.
Keywords: Colloidal gold; Rare Earth particles;silver staining;
目录
1 引言 1
1.1 免疫检测技术 1
1.2几种常用免疫方法 1
1.2.1 酶联免疫诊断技术 1
1.2.2 放射免疫分析 2
1.2.3 化学发光免疫分析 2
1.3 胶体Au标记技术 2
1.3.1 胶体Au的结构与特性 2
1.3.2 胶体Au标记蛋白质的原理 3
1.3.3 胶体Au的制备 3
1.3.4 胶体Au渗滤技术 4
1.4 稀土纳米材料及应用 4
1.4.1 稀土材料的介绍 4
1.4.2 稀土材料的应用 5
1.4.3 纳米稀土的制备 5
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