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本文作者用葡萄球菌A蛋白-辣根过氧化物酶(SPA-HRP)结合物间接免疫染色技术,检测石蜡包埋组织切片中的细胞内免疫球蛋白及其他抗原,并与过氧化物酶-抗过氧化物酶复合物(PAP)、“标记的抗原”和过氧化物酶标记的抗体夹心法等技术作比较.用过碘酸钠法将A蛋白与HRP结合,酶与A蛋白的用量比例为2:1(10毫克HRP∶5毫克SPA).SPA-HRP间接酶染色二步法程序是:第一步用特异性抗血清处理,第二步再用SPA-HRP处理.脱蜡的切片用含0.5%H_2O_2的甲醇处理5分钟,以阻断粒细胞、单核巨噬细咆和红细胞的内源性过氧化物酶活性.每张切片经pH7.6的0.05M Tfis-HCl冲洗后,各用一种抗血清——兔抗人,α,γ,μ,δ,重
The authors used the staphylococcal protein A-horseradish peroxidase (SPA-HRP) conjugate indirect immunostaining technique to detect intracellular immunoglobulins and other antigens in paraffin-embedded tissue sections and compared them with peroxidase- Anti-peroxidase complex (PAP), labeled antigen and peroxidase-labeled antibody sandwich method for comparison.A sodium salt method with protein A and HRP, enzyme and protein A (10 mg HRP: 5 mg SPA) .SPA-HRP indirect enzyme staining two-step procedure is: the first step with specific antiserum treatment, the second step with SPA-HRP treatment. Wax sections were treated with methanol containing 0.5% H 2 O 2 for 5 min to block the endogenous peroxidase activity of granulocytes, mononuclear macrophages and erythrocytes.Each slice was incubated with 0.05 M Tfis- After washing with HCl, each was treated with an antiserum - rabbit anti-human, α, γ, μ, δ, weight