摘要：本文用化学还原法制备RE微粒、纳米银微粒，研究了纳米银标记二抗的pH、二抗标记量、NC膜的预处理、一抗的浓度、封闭剂的种类、银染液浓度、RE与Ag的混合比例对银染效果的影响，进行了检出限的测定。采用紫外-可见分光光度计（UV-Vis）对纳米银、Ag-二抗、RE微粒、RE-Ag、RE-Ag-二抗进行表征。用Zeta电位仪分析了纳米银的电位。结果表明：硝酸纤文素膜进过预处理后，银染效果好；纳米银标记二抗的合适pH为7.4；加入二抗最少量5µg时，Ag-二抗复合物稳定存在；在一抗浓度为5μg/mL、银染液浓度为1.82×10-3 mol/L硝酸银和18.2×10-3 mol/L对苯二酚等体积混合时，银染效果较好。La 、Sm 、Ce、 Nd 用于与纳米银混合形成RE-Ag-二抗，银染效果好；在RE:Ag（RE：La、Ce、Sm、Nd）分别为4:6、3:7、2:8、1:9时，灰度值较大，其中La-Ag微粒的银染效果最好，检出限最低，为0.1fg/mL。本文创新之处：用稀土取代部分纳米银提高试剂盒的灵敏度。35855
毕业论文关键词：RE微粒；纳米银；银染
  Preparation of Kit for Detecting Concentration of TNNI3K
Using RE-Ag Filtration Assay
Abstract：ThisThe prepared by chemical reduction re particles, nano silver particles, the nano silver labeled pH second antibody and secondary antibody labeled amount, NC membrane pre processing, antibody concentration, sealant species, silver staining solution concentration, re and Ag mixing ratio on the effects of silver staining effect, the detection limits of the assay. UV VIS spectrophotometer (UV-Vis) was used to characterize the anti two, silver nanoparticles RE-Ag, RE-Ag- second antibody.. The potential of silver nanoparticles was analyzed by Zeta potential.. The results showed that the cellulose nitrate membrane through pretreatment, Silver stain effect is good; Nano silver mark second antibody suitable pH is 7.4 ;adding that the second anti amount for 5 µ g and Ag secondary antibody complexes are stable; in a antibody concentration is 5 u g / ml, silver dye concentration to 1.82×10-3 mol / L AgNO3 and    concentration for the mixed volume 18.2×10-3 mol / L hydroquinone etc., gray values of the best. For LA, SM, CE, Nd and nano silver mixture formation RE-Ag- second antibody, silver staining effect is good; in RE:Ag (re: La, CE, SM, nd) were 4:6, 3:7, 2:8 And 1:9, gray value is larger, the La-Ag particles of the silver dyeing effect is best, the minimum detection limit for 0.1fg/mL. The innovation of this paper is that the sensitivity of the rare earth to the improvement of the reagent boxes is used in the part of the rare earth..
Key Words: RE particles;silver nanoparticles; Silver staining
目录
1  引言    1
 1.1  诊断试剂盒的研究现状    1
 1.2  RE微粒的应用及制备方法    2
  1.2.1  RE在生物检测中的应用    2
  1.2.2  RE微粒的制备    2
 1.3  免疫渗滤法    3
 1.4  纳米银标记技术    4
  1.4.1  纳米银的特性    4
  1.4.2  纳米银的制备    4
  1.4.3  纳米银标记蛋白质的原理    5
 1.5  免疫检测技术    5
 1.6  研究内容和意义    6
  1.6.1  研究内容    6
  1.6.2  研究意义    6
2  实验部分    8
 2.1  实验材料与试剂    8
  2.1.1  实验材料    8
  2.1.2  实验试剂    8
 2.2  实验仪器    9
 2.3  纳米银和稀土微粒的制备    9 RE-Ag渗滤法制备TNNI3K浓度的检测试剂盒:http://www.youerw.com/huaxue/lunwen_34019.html