Antigen retrieval, blocking, detection and visualisation systems in immunohistochemistry: a review and practical
Anthony Warford1, Hameed Akbar1, Deise Riberio1
1University of Westminster, 115 New Cavendish Street, London W1W 6UW, United Kingdom.
Abstract:
To achieve specificity and sensitivity using immunohistochemistry it is necessary to combine the application of validated primary antibodies with optimised pre-treatment, detection and visualisation steps. The influence of these surrounding procedures is reviewed. A practical evaluation of tyramide signal amplification and rolling circle amplification detection methods is provided in which formalin fixed paraffin embedded sections of adenocarcinomas of breast, colon and lung together with squamous metaplasia of lung were immunostained with CD20 and CK19 primary antibodies. The results indicate that the detection systems are of comparable sensitivity and specificity.
