Abstract
Immunofluorescence (IF) of tumor tissues has become a key tool in the study of cancer. With a wide variety of formalin-fixed paraffin-embedded (FFPE) preserved tissues available, there are possibilities to assess large cohorts using archived tissue which may have archived associated clinical outcomes. Although best practice guidelines for the assessment of tissues have been published, a standardized method for immunofluorescence of FFPE tumor tissues is elusive. Here we provide a protocol for using classical secondary fluorescent antibodies that bind directly to the primary antibody of interest. This protocol can easily be adapted to use several primary antibodies, of different species, with unique secondary fluorophores that correspond to each species of origin. It can also be adapted for cyclic amplification-based immunofluorescence of FFPE tissues. We aim to provide a beginner-friendly and highly accessible method for immunofluorescence of FFPE-embedded tissues, hoping to enable more laboratories to take on this highly informative technique and empower them to begin IF analysis in their own tissues of interest.
Original language | English |
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Pages (from-to) | 169-181 |
Number of pages | 13 |
Journal | Methods in Molecular Biology |
Volume | 2508 |
DOIs | |
Publication status | Published - 2022 |
Bibliographical note
Publisher Copyright:© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
ASJC Scopus Subject Areas
- Molecular Biology
- Genetics
PubMed: MeSH publication types
- Journal Article