VISUALIZATION OF ANTIGENIC PROTEINS ON WESTERN BLOTS
VISUALIZATION OF ANTIGENIC PROTEINS ON WESTERN BLOTS
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DOI：
10.1016/0003-2697(84)90321-x
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发表时间：
1984-01-01
影响因子：
2.9
通讯作者：
DIMOND, RL
中科院分区：
文献类型：
--
作者：
KNECHT, DA;DIMOND, RL
建立了一种新的检测硝酸纤维素纸上蛋白质结合抗体的方法。该方法快速、灵敏，不需要放射性探针。转移到硝酸纤维素纸上的蛋白质首先与一抗反应，然后与碱性磷酸酶偶联的二抗反应。然后使用组织化学磷酸酶染色剂5-溴-4-氯-3-吲哚磷酸盐浸渍的琼脂凝胶观察磷酸酶活性。抗原抗体复合物在硝酸纤维素纸和琼脂覆盖凝胶上都会产生尖锐的、永久的蓝色染色条带。该程序允许检测含有少于20 ng蛋白质的条带。
A new technique for the detection of antibodies bound to proteins blotted onto nitrocellulose paper was developed. The method is rapid, sensitive and does not require radioactive probes. Proteins transferred to nitrocellulose paper are first reacted with primary antibody followed by reaction with an alkaline phosphatase conjugated second antibody. The phosphatase activity is then visualized using an agar gel impregnated with the histochemical phosphatase stain 5-bromo-4-chloro-3-indolyl phosphate. Antigen-antibody complexes give rise to sharp, permanent blue stained bands both on the nitrocellulose paper and in the agar overlay gel. This procedure allows detection of bands containing less than 20 ng of protein.