异戊烯基腺嘌呤核苷(iPA)的间接酶联免疫吸附测定法

Establishment of an Indirect Enzyme-linked Immunosorbent Assay for Isopentenyl Adenosine

以iPA－BSA为免疫抗原制备了兔抗血清，与其它iPA类似物的交叉反应率均低于5％，抗体－抗原亲和常数为6．26×106L·mol(-1)。利用此抗体与生物素－亲和素标记（ABC）系统建立了iPA的间接酶联免疫吸附测定法，检测线性范围为0．8～1000ng·mL(-1)（2．4～5693pmol．mL(-1)，板内误差CV＜5％，板间误差CV＜10％，样品平均回收率为87．3％。经平行试验、稀释试验以及对棉花根系伤流液、侧根组织和叶片中iPA含量的测定，表明所建立的ELISA及样品提取方法是可靠的。

The indirect enzyme-linked immunosorbent assay for isopentenyl adenosine (iPA) was established using rabbit antiserum against iPA-BSA and antirabbit IgG-BiotinAvidin-HRP labeling system. Association constant between antigen and antiserum was 6.26×106L·mol(-1) and all the cross reactivity value of antiserum with iPA similar compound were lower than 5%. The linear assay range of the method was 0.8~1000 ng·mL(-1).Variation within plate was less than 5%, and variation among plates was less than 10%.Average recovery rate of added standard in sample was 87. 3%. Some experiments was operated for quality control in practical assays. The contents of iPA in root bleedings, lateral roots and leaves of cotton plant were measured using this method.