Microwave oven-based technique for immunofluorescent staining of paraffin-embedded tissues.
Immunohistochemical analysis of formalin-fixed paraffin-embedded tissues can be challenging due to potential modifications of protein structure by exposure to formalin. Heat-induced antigen retrieval techniques can reverse reactions between formalin and proteins that block antibody recognition. Interactions between antibodies and antigens are further enhanced by microwave irradiation, which has simplified immunohistochemical staining protocols. In this report, we modify a technique for antigen retrieval and immunofluorescent staining of formalin-fixed paraffin-embedded tissues by showing that it works well with several antibodies and buffers. This microwave-assisted method for antigen retrieval and immunofluorescent staining eliminates the need for blocking reagents and extended washes, which greatly simplifies the protocol allowing one to complete the analysis in less than 3 h.
Long
Delwin J
DJ
Section of Pediatric Endocrinology and Metabolism, Baylor College of Medicine/Texas Children's Hospital, 1102 Bates St. Suite 550, Houston, TX, 77030, USA. DJLong@bcm.tmc.edu
Buggs
Colleen
C
D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D005260:Female; D005455:Fluorescent Antibody Technique; D005557:Formaldehyde; D007150:Immunohistochemistry; D051379:Mice; D008872:Microwaves; D016612:Paraffin Embedding; D010902:Pituitary Gland / Q000166:cytology; D013194:Staining and Labeling / Q000295:instrumentation* / Q000379:methods*; D016707:Tissue Fixation; D014599:Uterus / Q000166:cytology
D005557:Formaldehyde