A modification of the unlabeled antibody enzyme method using heterologous antisera for the light microscopic and ultrastructural localization of insulin, glucagon and growth hormone.
A modification of the unlabeled antibody enzyme method using heterologous antisera for the light microscopic and ultrastructural localization of insulin, glucagon and growth hormone.
复制标题
使用异源抗血清对未标记抗体酶法进行改进，用于胰岛素、胰高血糖素和生长激素的光显微镜和超微结构定位。
DOI：
10.1177/23.9.1176760
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发表时间：
1975
期刊：
影响因子：
--
通讯作者：
L. S. Van Orden
中科院分区：
文献类型：
--
作者：
S. Erlandsen;J. Parsons;J. P. Burke;J. Redick;D. V. van Orden;L. S. Van Orden
以胰岛素、胰高血糖素和生长激素的定位为模型系统，在光镜和电镜下研究了无标记抗体酶法中使用同源抗血清（一抗血清和同一物种中培养的过氧化物酶-抗过氧化物酶（PAP）复合物）的要求。用绵羊或山羊抗兔γ -球蛋白（S-ARgammaG或G-ARgammaG）偶联兔过氧化物酶-抗过氧化物酶复合物与豚鼠胰岛素抗血清（GP-AIS）或胰高血糖素抗血清（GP-AGS）或猴子大鼠生长激素抗血清（M-ARGH），观察到三种抗原的最佳免疫细胞化学染色。免疫电泳和放射免疫分析证实了S-ARgammaG或G-ARgammaG与这些初级抗血清中的免疫球蛋白的交叉反应性。S-ARgammaG与GP-AIS和M-ARGH分别与兔抗豚鼠γ -球蛋白抗血清和山羊抗猴γ -球蛋白抗血清反应时产生的沉淀弧相似。放射免疫分析结果显示，当S-ARgammaG用于免疫细胞化学时，与同源抗血清对照相比，6-10%的免疫沉淀获得了良好的染色定位。因此，只要偶联抗血清与原抗血清和PAP复合物的免疫球蛋白具有交叉反应性，异种抗血清（原抗血清和PAP复合物）可用于无标记抗体酶法。
The requirement of using homologous antisera (primary antiserum and peroxidase-antiperoxidase (PAP) complex raised in the same species) in the unlabeled antibody enzyme method has been investigated at the light and electron microscopic level using the localization of insulin, glucagon and growth hormone as model systems. Optimum immunocytochemical staining for all three antigens was observed when sheep or goat antirabbit gamma-globulin (S-ARgammaG or G-ARgammaG) were used to couple rabbit peroxidase-antiperoxidase complex with either guinea pig antisera to insulin (GP-AIS) or glucagon (GP-AGS), or monkey antisera to rat growth hormone (M-ARGH). The cross-reactivity between S-ARgammaG or G-ARgammaG and immunoglobulins in these primary antisera were substantiated by immunoelectrophoresis and radioimmunoassay. S-ARgammaG was shown to produce precipitation arcs with GP-AIS and M-ARGH that were similar to those seen when the latter were reacted with rabbit antiguinea pig gamma-globulin antiserum and goat antimonkey gamma-globulin antiserum, respectively. Radioimmunoassay results revealed that immunoprecipitation of 6-10% as compared to homologous antisera controls yielded excellent staining localization when S-ARgammaG was used for immunocytochemistry. Thus, heterologous antisera (primary antiserum and PAP complex raised in different species) may be used in the unlabeled antibody enzyme method as long as the coupling antiserum shows cross-reactivity with immunoglobulins of the primary antiserum and the PAP complex.