By Solheim J.C.
Well-recognized and cutting edge experimentalists aspect their state of the art tools for learning the antigen processing and presentation. Drawing on services from biochemistry, mobilephone biology, and immunology, they describe step by step tools designed to query 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 serious for experimental good fortune which are usually 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 useful instruments that may expand the questions that may be requested, and successfully be replied, bearing on antigen processing/presentation.
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Additional info for Antigen Processing and Presentation Protocols (Methods in Molecular Biology Vol 156)
Luckey, C. , King, G. , Marto, J. , Maier, B. , Crotzer, V. , et al. (1998) Proteasomes can either generate or destroy MHC class I epitopes: evidence for nonproteasomal epitope generation in the cytosol. J. Immunol. 161, 112–121. 33. , McMaster, J. , and Ploegh, H. L. (1998) A proteolytic system that compensates for loss of proteasome function. Nature 392, 618–622. 34. , and Niedermann, G. (1999) Giant protease with potential to substitute for some functions of the proteasome. Science 283, 978–981.
The introduction of proteasome inhibitors to cellular studies enabled the demonstration of the dominant role of this protease in cellular protein turnover and its involvement in the generation of class I ligands (6). Currently, there are four kinds of commonly used proteasome inhibitors, all of which, through different mechanisms, base their activity on the modification of the gamma oxygen (Oγ) on the N-terminal, active residue of Thr in one or more of the catalytic β-subunits (for a review on the mechanisms of the inhibitors, see ref.
Stock solutions of inhibitors as outlined in Table 2. 3. 1. Ruling Out Peptide Contamination Any study of the presentation of exogenous Ags on MHC-I must rule out peptide contamination in the Ag preparation. Despite attempts to remove peptide from Ag preparations by gel filtration, centrifugation, or other methods, it is possible that peptide contamination that is undetectable biochemically may still be present. Such peptide contamination will dramatically affect the results, and may be misleading.
Antigen Processing and Presentation Protocols (Methods in Molecular Biology Vol 156) by Solheim J.C.