Abstract
Major histocompatibility complex (MHC) class I molecules function to present pathogen-derived peptides to cytotoxic T cells or act as ligands for Natural Killer cells, thus alerting the immune system to the presence of invading pathogens. Furthermore MHC class I molecules can be strongly associated with autoimmune diseases. Therefore understanding not only the biosynthesis and the degradation pathways of MHC class I molecules has become important in determining their role in pathogen and autoimmune-related diseases. Here we describe how using epitope-tagged MHC class I molecules can aid in the analysis of MHC class I molecule biosynthesis and degradation and also complement studies using conventional conformationally specific antibodies. Coupled together with pharmacological manipulation which can target both biosynthetic and degradative pathways, this offers a powerful tool in analyzing MHC class I molecules.
Original language | English |
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Pages (from-to) | 93-108 |
Number of pages | 16 |
Journal | Methods in Molecular Biology |
Volume | 960 |
DOIs | |
Publication status | Published - 2013 |
Externally published | Yes |
Keywords
- Alleles
- Electrophoresis, Polyacrylamide Gel
- Epitopes/immunology
- Flow Cytometry
- HEK293 Cells
- HeLa Cells
- Histocompatibility Antigens Class I/biosynthesis
- Humans
- Immunoblotting
- Immunoprecipitation
- Proteolysis/drug effects