Journal
JOURNAL OF MOLECULAR HISTOLOGY
Volume 39, Issue 1, Pages 1-4Publisher
SPRINGER
DOI: 10.1007/s10735-007-9093-6
Keywords
microwave oven; immunofluorescence; antigen retrieval; pituitary gland; uterus; insulin receptor; insulin-like growth factor receptor; adipose
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Funding
- NIDDK NIH HHS [DK069518, L40 DK072905-02, K08 DK069518] Funding Source: Medline
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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.
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