ULTRASTRUCTURAL IMMUNOCYTOCHEMISTRY WITH UNLABELED ANTIBODIES AND THE PEROXIDASE-ANTIPEROXIDASE COMPLEX A TECHNIQUE MORE SENSITIVE THAN RADIOIMMUNOASSAY
ULTRASTRUCTURAL IMMUNOCYTOCHEMISTRY WITH UNLABELED ANTIBODIES AND THE PEROXIDASE-ANTIPEROXIDASE COMPLEX A TECHNIQUE MORE SENSITIVE THAN RADIOIMMUNOASSAY
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使用未标记抗体和过氧化物酶-抗过氧化物酶复合物的超结构免疫细胞化学是一种比放射免疫测定更灵敏的技术
DOI：
10.1177/21.9.825
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发表时间：
1973
影响因子：
3.2
通讯作者：
L. Sternberger
中科院分区：
文献类型：
--
作者：
G. C. Moriarty;C. Moriarty;L. Sternberger
用17- 39 ACTH和1- 39 ACTH抗血清，用放射免疫分析和电镜免疫细胞化学技术，绘制滴定曲线。后一种方法采用未标记抗体过氧化物酶-抗过氧化物酶复合物免疫细胞化学技术对正常大鼠垂体中间叶进行染色。用放射免疫分析法测定17- 39 ACTH抗血清的质量较差。其滴定曲线显示出平坦的斜率，并且其不结合超过1：30稀释度的显著量的标记抗原。然而，在高达1：1，500的稀释度下用该抗血清检测到免疫细胞化学染色。经放射免疫分析，1- 39 ACTH抗血清质量较好。在1：5，000的稀释度下，其结合45%的标记抗原。免疫细胞化学染色强度在1：5，000稀释度下接近最大值，并逐渐降低至1：16，000的极限值。然而，当在抗血清中的孵育时间从3分钟增加到与放射免疫测定的孵育时间（48小时）相匹配时，在稀释度高达1：512，000时达到最大染色，其中在放射免疫测定中仅结合痕量的标记抗原。它的结论是，未标记的抗体过氧化物酶-抗过氧化物酶复合物免疫细胞化学技术是足够敏感的检测抗体的低滴度和/或亲和力，这是没有检测到的放射免疫分析法血清。该技术为灵敏的测定系统提供了很大的希望。
Titration curves were developed with antisera to 17-39ACTH (adrenocorticotropin) and 1-39ACTH with the techniques of radioimmunoassay and electron microscopic immunocytochemistry. For the latter method, the unlabeled antibody peroxidase-antiperoxidase complex immunocytochemical technique was used to stain normal rat pituitary intermediate lobes. By radioimmunoassay standards, the 17-39ACTH antiserum was of poor quality. Its titration curve exhibited a flat slope and it did not bind a significant amount of labeled antigen beyond a 1: 30 dilution. However, immunocytochemical staining was detected with this antiserum at dilutions as high as 1:1,500. The antiserum to 1-39ACTH was of better quality by radioimmunoassay standards. It bound 45% of the labeled antigen at a dilution of 1:5,000. Immunocytochemical staining intensity was nearly maximal at a 1:5,000 dilution and decreased progressively to a limiting value at 1:16,000. However, when incubation times in the antisera were increased from 3 min to match those of the radioimmunoassay (48 hr) maximal staining was achieved at dilutions as great as 1:512,000 where only trace amounts of the labeled antigen were bound in the radioimmunoassay. It was concluded that the unlabeled antibody peroxidase-antiperoxidase complex immunocytochemical technique was sensitive enough to detect antibodies in sera of low titer and/or avidity which are not detected by a radioimmunoassay. The technique holds great promise for a sensitive assay system.