Application of the avidin-biotin-peroxidase complex (ABC) method to the light microscopic localization of pituitary hormones.
Application of the avidin-biotin-peroxidase complex (ABC) method to the light microscopic localization of pituitary hormones.
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DOI：
10.1177/30.7.6286753
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发表时间：
1982-07
期刊：
影响因子：
--
通讯作者：
G. Childs;G. Unabia
中科院分区：
文献类型：
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作者：
G. Childs;G. Unabia
抗生物素蛋白-生物素-过氧化物酶复合物（ABC）法应用于半薄（0.5-1微米）塑料包埋切片的完整雄性大鼠垂体与使用技术先前开发的过氧化物酶-抗过氧化物酶复合物（PAP）的方法。促肾上腺皮质激素（ACTH）、促甲状腺激素（TSH）、促黄体生成素（LH）和促卵泡激素（FSH）染色更清晰、更可靠、更有效。ABC法允许使用与PAP法相同的高稀释度的一级抗血清。孵育时间缩短至PAP染色时间的三分之一。此外，如果孵育时间与PAP方法所用的孵育时间相匹配（24-48小时），则抗血清可进一步稀释2- 4倍。这增强了特异性染色，并允许使用与放射免疫测定中使用的稀释液相似的稀释液。与Hsu和Raine（J Histochem Cytochem 29：1349，1981）一致，ABC法在PAP复合物法最佳稀释的一抗中仅孵育1-4小时后就产生染色。然而，染色很弱，细胞计数表明它仅限于储存最多激素的特定细胞群的部分。结果表明，最佳染色时间为12-16小时，且ABC法能使FSH反应稳定，提高了其精密度和可靠性。
The avidin-biotin-peroxidase complex (ABC) method was applied to semithin (0.5-1 micron) plastic-embedded sections of intact male rat pituitaries with the use of techniques previously developed for the peroxidase-antiperoxidase complex (PAP) method. Stains for adrenocorticotropin (ACTH), thyroid stimulating hormone (TSH), luteinizing hormone (LH), and follicle stimulating hormone (FSH) were cleaner, more reliable, and more efficient. The ABC method allowed the use of the same high dilutions of primary antisera used with the PAP method. Incubation time was cut to a third of the time used for the PAP stain. Furthermore, if the incubation time matched that used with the PAP method, (24-48 hr), the antisera could be diluted 2- to 4-fold further. This enhanced specific staining and allowed the use of dilutions similar to those used in the radioimmunoassay. In agreement with Hsu and Raine (J Histochem Cytochem 29:1349, 1981), the ABC method produced staining after only a 1-4 hr incubation in primary antibody that was diluted optimally for the PAP complex method. The stain was weak, however, and cell counts showed that it was restricted to the fraction of the specific cell population which stored the most hormone. Our tests showed that the most convenient incubation times for optimal staining were 12-16 hr. Furthermore, the ABC method appeared to stabilize greatly the reaction for FSH and thus improved its precision and reliability.