一种用于检测高效液相色谱法纯化的植物提取物中飞摩尔级异戊烯基腺嘌呤和异戊烯基腺苷的抗生物素蛋白-生物素固相酶联免疫吸附测定法。
An avidin-biotin solid phase ELISA for femtomole isopentenyladenine and isopentenyladenosine measurements in HPLC purified plant extracts.
作者信息
Sotta B, Pilate G, Pelese F, Sabbagh I, Bonnet M, Maldiney R
机构信息
Laboratoire de Physiologie Végétale, T 53, 5ème étage, Université P et M Curie, 4 place Jussieu, 75252 Paris Cedex 05, France.
出版信息
Plant Physiol. 1987 Jul;84(3):571-3. doi: 10.1104/pp.84.3.571.
Abstract
A solid phase enzyme immunoassay was developed for isopentenyladenine (iP) and isopentenyladenosine (iPA) quantitation in HPLC purified plant extracts. It was performed on antigen-coated microtitration plates on which bound antibodies were indirectly labeled by the means of a biotinylated goat anti-rabbit antibody and an avidin-alkaline phosphatase conjugate. Less than 3 femtomoles of iP or iPA were easily detected and the measuring range extended from 3 femtomole to 1 picomole. The reproducibility has been tested and intra- and interassay variations did not exceed 5.0%. The specificity of iPA antibodies was good, as determined by cross-reactivity measurements with other adenylic compounds. The specificity of the measurements for iP and iPA was demonstrated by analysis of the immunoreactivity of fractions obtained by HPLC separation of a methanolic tobacco leaf extract.
摘要
开发了一种固相酶免疫测定法，用于对HPLC纯化的植物提取物中的异戊烯基腺嘌呤（iP）和异戊烯基腺苷（iPA）进行定量。该测定法在包被抗原的微量滴定板上进行，结合的抗体通过生物素化山羊抗兔抗体和抗生物素蛋白-碱性磷酸酶缀合物间接标记。易于检测到少于3飞摩尔的iP或iPA，测量范围从3飞摩尔扩展到1皮摩尔。已测试了重现性，批内和批间变异不超过5.0%。通过与其他腺苷酸化合物的交叉反应测量确定，iPA抗体的特异性良好。通过对甲醇提取的烟草叶提取物进行HPLC分离得到的级分的免疫反应性分析，证明了对iP和iPA测量的特异性。
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