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免疫组织化学技术在国内已广泛开展。但由于国内抗血清来源困难,尤其是第一抗血清(一抗)的紧缺,给这项工作的普及设置了障碍。本实验室在开展消化道神经肽免疫组化工作中,将用过的抗血清做回收再用,收到较好的效果。介绍如下:1.抗血清:一抗本实验选用四种抗血清:血管活性肠肽(VIP)、物质P(sp)、生长抑制(SOM)和亮啡肽(LNK)抗血清(Immuno.Nuclean Comp.)。工作浓度均为1∶6000。二抗(羊抗兔血清)及PAP来自上海医科大学病理科。二抗工作浓度为1∶40;PAP为1∶80。2.取材:雄性大白鼠两只,体重约200克。4%戊巴比妥钠腹腔注射麻醉(3-4 mg/100 g)。
Immunohistochemistry has been widely carried out in China. However, due to the difficulties of sources of domestic antisera, especially the shortage of the first antiserum (primary antibody), this has set an obstacle for the popularization of this work. The laboratory in carrying out the work of immunohistochemistry of the digestive tract neuropeptide, the used antisera to do recycling, received good results. The introduction is as follows: 1. Antiserum: primary antibody Four kinds of antisera were selected for this experiment: vasoactive intestinal peptide (VIP), substance P (sp), growth inhibition (SOM) and leupeptin (LNK) antiserum Comp.). Working concentrations are 1: 6000. Secondary antibody (goat anti-rabbit serum) and PAP from the Department of Pathology, Shanghai Medical University. Secondary antibody working concentration of 1:40; PAP of 1:80.2. Draws: two male rats, weighing about 200 grams. 4% pentobarbital sodium intraperitoneal anesthesia (3-4 mg / 100 g).