使用免疫荧光染色评估福尔马林固定石蜡包埋组织切片上 GD2 的方案。

Protocol for assessing GD2 on formalin-fixed paraffin-embedded tissue sections using immunofluorescence staining.

机构信息

Department of Pediatric Hematology and Oncology, University Children's Hospital Muenster, Muenster, Germany; Princess Máxima Center for Pediatric Oncology, Utrecht, the Netherlands.

Department of Pediatric Hematology and Oncology, University Children's Hospital Muenster, Muenster, Germany.

出版信息

STAR Protoc. 2024 Sep 20;5(3):103199. doi: 10.1016/j.xpro.2024.103199. Epub 2024 Jul 22.

DOI:10.1016/j.xpro.2024.103199

PMID:39046881

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11321291/

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..

摘要

由于 GD2 的糖脂结构及其膜锚定，肿瘤活检标本（尤其是石蜡包埋组织）中 GD2 的检测一直具有挑战性。在此，我们提出了一种可靠评估福尔马林固定石蜡包埋（FFPE）组织中 GD2 的免疫荧光方案。我们描述了使用 Tris-EDTA 缓冲液进行抗原修复以及使用未缀合的抗 GD2 抗体（克隆 14.G2a）和辣根过氧化物酶（HRP）缀合的二抗进行染色的步骤。然后，我们详细介绍了使用酪胺信号放大技术进行信号放大的程序。有关此方案使用和执行的完整详细信息，请参阅 Fischer-Riepe 等人的研究。