By Solheim J.C.
Well-recognized and cutting edge experimentalists element their state-of-the-art tools for learning the antigen processing and presentation. Drawing on services from biochemistry, phone biology, and immunology, they describe step by step tools designed to question how MHC-binding peptides are generated, to check how peptides are dropped at MHC molecules, to research MHC peptide-binding styles, and to assay the T-cell reaction to the MHC/peptide advanced. Emphasis is given these technical steps severe for experimental luck which are frequently passed over from tools released within the fundamental literature. Eminently obtainable and state of the art, Antigen Processing and Presentation Protocols offers either new and skilled investigators with hugely sensible instruments that may expand the questions that may be requested, and successfully be spoke back, touching on antigen processing/presentation.
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Extra resources for Antigen Processing and Presentation Protocols (Methods in Molecular Biology Vol 156)
There are two chief methods of accomplishing this. The most widely used strategy entails transient expression of the substrate, which is achieved by either viral infection of the target cells or by loading of the purified protein into the cytosol; alternatively, cells constitutively expressing a target Ag are acid-stripped to remove existing complexes, and the effect of proteasome inhibitors on regeneration of peptide–class I complexes is determined. As alluded to above, one must exercise caution when interpreting results from any experiment using proteasome inhibitors.
Chem. 273, 6373–6379. Yellen-Shaw, A. J. C. (1997) Regulation of class I-restricted epitope processing by local or distal flanking sequence. J. Immunol. 158, 1727–1733. , Antón, L. , Snyder, H. , Bennink, J. , and Yewdell, J. W. Generation of MHC class I-associated peptides is only partially inhibited by proteasome inhibitors. Involvement of nonproteasomal proteases in antigen processing? J. Immunol. 159, 554–564. , and Townsend, A. (1997) Proteasome-specific inhibitor lactacystin blocks presentation of cytotoxic T lymphocyte epitopes in human and murine cells.
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