Peterman T K, Siedow J N
Arch Biochem Biophys. 1985 May 1;238(2):476-83. doi: 10.1016/0003-9861(85)90190-0.
An affinity-purified polyclonal antibody against soybean seed lipoxygenase-2 was prepared and used to characterize the immunological relatedness of lipoxygenase isozymes 1 and 2 and lipoxygenases from soybean seedling roots, hypocotyls, leaves, and cotyledons. All soybean lipoxygenases tested cross-reacted with the anti-lipoxygenase-2. Cross-reactivity of seed-derived lipoxygenases was evidenced by formation of a line of identity in double-diffusion tests, by positive results on an immunoblot, and by antibody precipitation of enzyme activity. Levels of anti-lipoxygenase-2, which inhibited lipoxygenase-2 activity, had no effect on lipoxygenase-1 activity. Root, hypocotyl, and leaf lipoxygenases did not form precipitation lines in double-diffusion tests but the anti-lipoxygenase-2 did inhibit and precipitate lipoxygenase activity from these sources as well as cross-react on immunoblots. All the cross-reactive lipoxygenases examined were found to have the same apparent molecular weight. Lipoxygenase activity found in soybean seedling roots, hypocotyls, leaves, and cotyledons is associated with proteins which are all immunologically related to the seed-derived enzymes.
制备了一种针对大豆种子脂氧合酶-2的亲和纯化多克隆抗体，并用于表征脂氧合酶同工酶1和2以及来自大豆幼苗根、下胚轴、叶片和子叶的脂氧合酶的免疫相关性。所有测试的大豆脂氧合酶都与抗脂氧合酶-2发生交叉反应。种子衍生的脂氧合酶的交叉反应性通过双向扩散试验中同一性线的形成、免疫印迹的阳性结果以及酶活性的抗体沉淀来证明。抑制脂氧合酶-2活性的抗脂氧合酶-2水平对脂氧合酶-1活性没有影响。根、下胚轴和叶片脂氧合酶在双向扩散试验中未形成沉淀线，但抗脂氧合酶-2确实抑制并沉淀了这些来源的脂氧合酶活性，并且在免疫印迹上发生交叉反应。所有检测的交叉反应性脂氧合酶都具有相同的表观分子量。在大豆幼苗根、下胚轴、叶片和子叶中发现的脂氧合酶活性与所有与种子衍生酶具有免疫相关性的蛋白质相关。
