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Published on: July 13, 2016
Protocol for assessing GD2 on formalin-fixed paraffin-embedded tissue sections using immunofluorescence staining
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.
Insights
This study introduces a new immunofluorescence protocol for accurately detecting disialoganglioside GD2 in formalin-fixed paraffin-embedded tumor tissues. The method enhances GD2 detection in challenging FFPE samples for improved diagnostic accuracy.
Area of Science:
- Oncology
- Immunohistochemistry
- Biomarker Detection
Background:
- Disialoganglioside GD2 detection on tumor biopsies, particularly in FFPE tissues, is difficult due to its glycolipid nature and membrane anchoring.
- Reliable GD2 assessment is crucial for certain cancer diagnostics and therapeutic strategies.
Purpose of the Study:
- To develop and present a robust immunofluorescence protocol for the reliable assessment of disialoganglioside GD2 in formalin-fixed paraffin-embedded (FFPE) tissues.
- To overcome the challenges associated with GD2 detection in FFPE samples.
Main Methods:
- The protocol involves antigen retrieval using Tris-EDTA buffer.
- Staining utilizes an unconjugated anti-GD2 antibody (clone 14.G2a) and a horse radish peroxidase (HRP)-conjugated secondary antibody.
- Tyramide signal amplification technique is employed for enhanced signal detection.
Main Results:
- The described immunofluorescence protocol enables reliable detection of disialoganglioside GD2 on FFPE tissues.
- The method addresses the inherent challenges of GD2 detection in fixed tissue samples.
Conclusions:
- This protocol provides a valuable tool for researchers and clinicians needing to assess GD2 expression in FFPE tumor biopsies.
- The optimized immunofluorescence method facilitates more accurate and consistent GD2 detection in challenging FFPE samples.
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.
