Protocol for assessing GD2 on formalin-fixed paraffin-embedded tissue sections using immunofluorescence staining
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Published on: July 13, 2016
Sareetha Kailayangiri1, Bianca Altvater1, Nicole Farwick2
1Department of Pediatric Hematology and Oncology, University Children's Hospital Muenster, Muenster, Germany; Princess Máxima Center for Pediatric Oncology, Utrecht, the Netherlands.
Abstract:
The detection of disialoganglioside GD2 on tumor biopsies, especially in paraffin-embedded tissues, has been challenging due to the glycolipid structure of GD2 and its membrane anchorage. Here, we present an immunofluorescence protocol for the reliable assessment of GD2 on formalin-fixed paraffin-embedded (FFPE) tissues. We describe steps for antigen retrieval with Tris-EDTA buffer and staining with unconjugated anti-GD2 antibody (clone 14.G2a) and horse radish peroxidase (HRP)-conjugated secondary antibody. We then detail procedures for signal amplification using the tyramide signal amplification technique. For complete details on the use and execution of this protocol, please refer to Fischer-Riepe et al.1.
