【摘要】 作者应用四种抗血清,在不同孵育条件下比较PAP、双桥PAP及ABC法的敏感性。双桥PAP法为一实用、敏感的免疫组化法。探讨了提高免疫组化染色强度的操作步骤:用延长第一抗体孵育时间(过夜),抗体稀释度提高5～10倍,第二、三步骤用提高孵育温度(37℃)来缩短孵育时间,可获得满意的染色效果。更多还原

【Abstract】 The sensitivity of PAP,double-bridgePAP and ABC mothods was compared,using anti-GFAP,anti-S-100 protein,anti-growth hormone and anti-ACTH.All pro-cedures were carried out under-three con-ditions: 1.in moist chamber;2.on hotplate; and 3. incubation of primary antibodies inmoist chamber,and the remainder proce-dures were completed on hotplate. The results reveal that ABC mothod isthe most sensitive one among the three,andthe next one is double-bridge PAP mothod.As far as amiprotein serum is concerned,ABC method is a little more sensitive thandouble-bridge PAP method,but 5 timesmore sensitive when antipolypeptide reagentis used. The prolongation of incubation time ofprimary antibodies may make more dilutedprimary antibody used,and save primaryantibody.The incubation time for linkingantibody,biotinylated antibody,PAP com-plex and ABC complex may be shortenedto 5 minutes (37℃) with satisfactory stain-ing.更多还原