Kaneko T, Fraser R A, Labedz T, Harvey S, Lafeber F P, Pang P K
Department of Physiology, University of Alberta, Edmonton, Canada.
Gen Comp Endocrinol. 1988 Feb;69(2):238-45. doi: 10.1016/0016-6480(88)90011-1.
Highly specific antisera were raised against hypocalcin, a 54-kDa glycoprotein purified from the corpuscles of Stannius (CS) of rainbow trout, Salmo gairdneri. The specificity of the antisera was determined by using Ouchterlony's double immunodiffusion test, radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), and immunocytochemistry. In the double immunodiffusion test, a single precipitin line was formed between the antisera and hypocalcin. Both RIA and ELISA studies showed that serial dilutions of hypocalcin, teleocalcin, and CS extracts produced dose-response curves, whereas rat FSH, chicken LH, bovine TSH, salmon calcitonin, bovine parathyroid hormone, and human angiotensins I and II failed to cross-react with the antiserum. Immunoreactive hypocalcin was demonstrated in the plasma of a teleost (flounder), but not of an elasmobranch (dogfish). In the immunocytochemistry, most of the gland cells showed strong immunoreaction with the antisera, whereas some cells displayed no immunoreactivity.
制备了针对低钙素的高度特异性抗血清,低钙素是一种从虹鳟(Salmo gairdneri)斯坦尼斯小体(CS)中纯化得到的54 kDa糖蛋白。通过免疫双扩散试验、放射免疫测定(RIA)、酶联免疫吸附测定(ELISA)和免疫细胞化学来确定抗血清的特异性。在免疫双扩散试验中,抗血清与低钙素之间形成了一条单一的沉淀线。RIA和ELISA研究均表明,低钙素、远钙素和CS提取物的系列稀释产生了剂量反应曲线,而大鼠促卵泡激素、鸡促黄体激素、牛促甲状腺激素、鲑鱼降钙素、牛甲状旁腺激素以及人血管紧张素I和II均未与抗血清发生交叉反应。在一种硬骨鱼(比目鱼)的血浆中检测到了免疫反应性低钙素,但在一种软骨鱼(角鲨)中未检测到。在免疫细胞化学中,大多数腺细胞与抗血清呈现强烈的免疫反应,而一些细胞则无免疫反应。