中国药理学通报Issue(3)：436-440,441,6.DOI:10.3969/j.issn.1001-1978.2015.03.028
一种简便、可靠的免疫组化双重标记新方法
A novel,convenient and reliable method for immunohistochemical double labeling
摘要
Abstract
Aim To establish a convenient, reliable and high- <br> contrast method of immunohistochemical double labeling, <br> which can be observed with ordinary optical microscopy, and the staining results can be preserved permanently. Method This double labeling method employed the single detection system, HRP conjugated second antibodies, and used the substrates of 3,3′-diaminobenzidine ( DAB) and Nickel-enhanced DAB re-spectively. When the first labeling was completed, HRP was to-tally inactivated by boiling for 4~5 min to prevent the cross-re-action in the second labeling. Results ( 1 ) Boiling for 5 min could completely inactivate HRP activity, whereas 5% H2 O2 treatment only reduced HRP activity, and residual enzyme activ-ity generated the cross-reaction between two labeling. Boiling for a few minutes did not damage positive DAB staining in the first labeling or tissue integrity. (2) There was a sharp contrast be- <br> tween DAB in brown and Nickel enhanced DAB in purple-blue, which also gives clean backgrounds;reversely applying the sub-strates will ruin the quality of double labeling. In addition, the above method allowed conventional mounting protocol using xy-lene and Permount, instead of a water-based mounting medium, thus the staining results could be permanently preserved. Con-clusion As the same HRP detection system is employed, this novel double labeling method is simple, convenient and reliable. The method would bear broad applications in basic and clinical research.关键词
免疫组化/辣根过氧化物酶/二氨基联苯胺/镍/方法/双重标记/交叉反应Key words
immunohistochemistry/horseradish peroxidase/3, 3′-diaminobenzidine/Nickel/method/double labeling/cross-reaction引用本文复制引用
崔白苹,高璀乡,熊存全,孙安阳..一种简便、可靠的免疫组化双重标记新方法[J].中国药理学通报,2015,(3):436-440,441,6.基金项目
国家自然科学基金资助项目(30470594,30772282) （30470594,30772282）
