Glazer E J, Ramachandran J, Basbaum A I
J Histochem Cytochem. 1984 Jul;32(7):778-82. doi: 10.1177/32.7.6203962.
Affinity-purified goat anti-rabbit immunoglobulin G (GAR) was conjugated with (3H)-propionyl succinimidate and used to localize substance P (SP), enkephalin (ENK), and serotonin immunoreactive sites in the spinal dorsal horn and medulla of the rat and cat. Autoradiographic localization was demonstrated on paraffin, frozen, Vibratome, and 2 micron plastic sections. The latter were obtained from radiolabeled Vibratome sections that were embedded in epoxy resin. The distribution of SP, ENK, and serotonin demonstrated by radioimmunocytochemistry was comparable to that observed on semiadjacent sections using peroxidase-antiperoxidase (PAP) immunocytochemistry. The autoradiograms, however, were generated using primary antibody concentrations up to five times more dilute than concentrations used for the PAP procedure. Indirect radioimmunocytochemistry using a (3H) anti-immunoglobulin G second antibody can be used to localize a variety of monoclonal and polyclonal antisera. It is quantifiable at the light microscopic level and can be potentially used with peroxidase histochemistry to double label immunoreactive structures at the ultrastructural level.
亲和纯化的山羊抗兔免疫球蛋白G（GAR）与（3H）-丙酰琥珀酰亚胺结合，用于定位大鼠和猫脊髓背角及延髓中P物质（SP）、脑啡肽（ENK）和5-羟色胺免疫反应位点。在石蜡切片、冰冻切片、振动切片机切片和2微米塑料切片上进行放射自显影定位。后者取自嵌入环氧树脂的放射性标记振动切片机切片。放射免疫细胞化学显示的SP、ENK和5-羟色胺的分布与使用过氧化物酶-抗过氧化物酶（PAP）免疫细胞化学在半相邻切片上观察到的分布相当。然而，放射自显影片是使用比PAP程序所用浓度稀释高达五倍的一抗浓度生成的。使用（3H）抗免疫球蛋白G二抗的间接放射免疫细胞化学可用于定位多种单克隆和多克隆抗血清。它在光学显微镜水平上是可量化的，并且有可能与过氧化物酶组织化学一起用于在超微结构水平上对免疫反应结构进行双重标记。
