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利用棋盘格实验直接筛选酶免疫测定的最佳包被抗原和抗体浓度 被引量:12
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作者 谭伟明 何素平 +6 位作者 张蕾 赵洪伟 赵静 李召虎 李学锋 王保民 qing x. li 《分析化学》 SCIE EI CAS CSCD 北大核心 2008年第9期1191-1195,共5页
以两个单克隆抗体及其相应的包被抗原为例,研究了棋盘格实验中包被抗原与抗体浓度组合对免疫检测中0孔吸光度值A0、IC50值和标准曲线斜率的影响。结果发现,A0和IC50值随包被抗原与抗体稀释倍数的增加而降低;斜率随抗体稀释倍数增加而降... 以两个单克隆抗体及其相应的包被抗原为例,研究了棋盘格实验中包被抗原与抗体浓度组合对免疫检测中0孔吸光度值A0、IC50值和标准曲线斜率的影响。结果发现,A0和IC50值随包被抗原与抗体稀释倍数的增加而降低;斜率随抗体稀释倍数增加而降低,而随包被抗原的稀释倍数增加表现出先增加后降低的趋势。在同一抗体浓度和不同包被抗原浓度的组合中,斜率最大值出现在使板吸附达到饱和的最大稀释倍数的包被抗原浓度附近。综合实验结果确认从棋盘格实验中直接筛选最佳包被抗原、抗体浓度组合的标准是:选择使板吸附达到饱和的最大包被抗原稀释倍数为最佳包被抗原浓度,在不同抗体浓度与已选择的最佳包被抗原浓度组合中使0孔吸光度值达1.0左右的抗体稀释倍数为最佳抗体浓度。 展开更多
关键词 酶联免疫吸附测定 棋盘格实验 方法优化
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Monoclonal Antibody-Based Enzyme Linked Immunosorbent Assay for the Analysis of Jasmonates in Plants 被引量:10
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作者 Aixing Deng Weiming Tan +5 位作者 Suping He Wei liu Tiegui Nan Zhaohu li Baomin Wang qing x. li 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2008年第8期1046-1052,共7页
Methyl jasmonate (MeJA) and its free-acid form, jasmonic acid (JA) are naturally occurring plant growth regulators widely distributed in higher plants. In order to improve the sensitivity for the analysis of MeJA ... Methyl jasmonate (MeJA) and its free-acid form, jasmonic acid (JA) are naturally occurring plant growth regulators widely distributed in higher plants. In order to improve the sensitivity for the analysis of MeJA at low levels in small amounts of plant samples, a monoclonal antibody (MAb) (designated as MAb 3E5D7C4B6) against MeJA was derived from a JA- bovine serum albumin (BSA) conjugate as an immunogen. The antibody belongs to the IgG1 subclass with a κ type light chain and has a dissociation constant of approximately 6.07 ×10^-9 M. MAb3E5D7C4B6 is very specific to MeJA. It was used to develop a direct competitive enzyme-linked immunosorbent assay (dcELISA), conventional and simplified indirect competitive ELISAs (icELISA). JA was derivatized into MeJA for the ELISA analysis. The IC50 value and detection range for MeJA were, respectively, 34 and 4-257 nglmL by the conventional icELISA, 21 and 3-226 nglmL by the simplified icELISA and 5.0 and 0.7-97.0 nglmL by the dcELISA. The dcELISA was more sensitive than either the conventional or simplified icELISA. The assays were used to measure the content of jasmonates as MeJA in tobacco leaves under drought stress or inoculated with tobacco mosaic virus and tomato leaves inoculated with tomato mosaic virus or Lirioinyza sativae Blanchard as compared with the corresponding healthy leaves. The increased jasmonates content indicated its role in response to the drought stress and pathogens. 展开更多
关键词 enzyme-linked immunosorbent assay IMMUNOASSAY methyl jasmonate plant growth regulators
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