DETECTION OF ANTIGENS ON NITROCELLULOSE PAPER IMMUNOBLOTS WITH MONOCLONAL-ANTIBODIES
DETECTION OF ANTIGENS ON NITROCELLULOSE PAPER IMMUNOBLOTS WITH MONOCLONAL-ANTIBODIES
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DOI：
10.1016/0003-2697(83)90245-2
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发表时间：
1983-01-01
影响因子：
2.9
通讯作者：
CHERWINSKI, HM
中科院分区：
文献类型：
--
作者：
DEBLAS, AL;CHERWINSKI, HM
描述了一种非常灵敏的检测硝酸纤维素纸免疫印迹上抗原抗体复合物的方法。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离蛋白抗原，然后将其电泳转移到硝酸纤维素片上（“Western印迹”）。将与硝酸纤维素纸结合的蛋白质抗原暴露于单克隆抗体，并通过免疫酶反应在纸上检测抗体-抗原复合物。该方法灵敏度的提高是由于（i）使用去污剂吐温20来阻断抗体与硝酸纤维素纸的非特异性结合，（ii）使用过氧化物酶-抗过氧化物酶（PAP）反应，以及（iii）增强磷酸盐缓冲液中二氨基联苯胺与镍和钴离子的反应产物。
A very sensitive method for the detection of antigen-antibody complexes on nitrocellulose paper immunoblots is described. The protein antigens are separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by their electrophoretic transfer onto a nitrocellulose sheet (“Western blot”). The protein antigens bound to the nitrocellulose paper are exposed to the monoclonal antibody and the antibody-antigen complexes are detected on the paper by an immunoenzymatic reaction. The improved sensitivity of this method is the result of (i) the use of the detergent Tween 20 in blocking the nonspecific binding of the antibodies to the nitrocellulose paper, (ii) the use of a peroxidase-antiperoxidase (PAP) reaction, and (iii) the intensification of the diaminobenzidine reaction product with nickel and cobalt ions in phosphate buffer.