Professional antigen-presenting cells such as dendritic cells (DCs) and macrophages internalize extracellular antigens, process them intracellularly, and present the resulting antigen-derived peptides in the context of MHC I or MHC II molecules. Since the intracellular routing of the antigen determines whether antigens are presented on MHC I or MHC II molecules, a profound analysis of the intracellular distribution of the internalized antigens is of high interest. Here, we describe an immuno fl uorescence protocol to monitor the intracellular routing of the model-antigen Ovalbumin in bone marrow-derived dendritic cells (BM-DCs). This protocol describes a procedure to stain such cells with antibodies against different endosomal markers, such as EEA1 and LAMP1, and can be easily adopted to other antigen-presenting cells or antigens. © Springer Science+Business Media, LLC 2013.
CITATION STYLE
Franken, L., Kurts, C., & Burgdorf, S. (2013). Monitoring the intracellular routing of internalized antigens by immuno fl uorescence microscopy. Methods in Molecular Biology, 960, 371–377. https://doi.org/10.1007/978-1-62703-218-6_27
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