By Stefan Tenzer, Tobias Hain, Hendrik Berger, Hansjörg Schild (auth.), Peter van Endert (eds.)
Antigen processing is a organic technique that prepares antigens for the presentation to important cells within the immune process known as T lymphocytes. In Antigen Processing: equipment and Protocols, specialist researchers within the box supply a accomplished set of protocols for learning presentation of antigens produced within the typical processing pathways for MHC classification I and sophistication II molecules. The chapters stick with chronology of intracellular processing occasions, finishing with acceptance of peptide-MHC complexes on the mobilephone floor via T lymphocytes. Written within the hugely profitable Methods in Molecular Biology™ sequence layout, chapters comprise introductions to their respective themes, lists of the mandatory fabrics and reagents, step by step, with ease reproducible laboratory protocols, and key pointers on troubleshooting and heading off identified pitfalls.
Authoritative and functional, Antigen Processing: tools and Protocols is designed for newbies and specialists attracted to learning antigen processing.
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Additional info for Antigen Processing: Methods and Protocols
Antigen Processing: Methods and Protocols, Methods in Molecular Biology, vol. 1007/978-1-62703-218-6_2, © Springer Science+Business Media, LLC 2013 15 16 K. Textoris-Taube et al. the presence of immunoproteasomes and PA28 (5–10) influence epitope liberation either positively or negatively. To study proteasomal antigen processing simple in vitro systems have been established that allow mimicking the in vivo situation with surprisingly high fidelity. These assays generally involve the processing of a synthetic polypeptide of 20–30 amino acid residues in length that harbor a known or a potential MHC class I epitope by purified 20S proteasomes, HPLC-separation of the generated peptides followed by MS analysis of the generated peptide products (11) (see Note 1).
You need an appropriate laser in your flow cytometer to excite your fluorescent dye and also detectors measuring the correct emission wavelength. 38 M. Basler and M. Groettrup 14. Do not count residual un-lysed erythrocytes. 15. It is useful to pulse female splenocytes with different concentrations of synthetic peptide (10−5 to 10−10 M) and use the pulsed cells as stimulators for hybridomas. Comparison of the stimulation of the titrated peptide probes with your samples will provide information regarding the magnitude of observed differences.
Nat Immunol 11:449–454 3. Stoltze L, Schirle M, Schwarz G et al (2000) Two new proteases in the MHC class I processing pathway. Nat Immunol 1:413–418 4. Saric T, Beninga J, Graef CI et al (2001) Major histocompatibility complex class I-presented antigenic peptides are degraded in cytosolic extracts primarily by thimet oligopeptidase. J Biol Chem 276:36474–36481 5. York IA, Mo AXY, Lemerise K et al (2003) The cytosolic endopeptidase, thimet oligopeptidase, destroys antigenic peptides and limits the extent of MHC class I antigen presentation.
Antigen Processing: Methods and Protocols by Stefan Tenzer, Tobias Hain, Hendrik Berger, Hansjörg Schild (auth.), Peter van Endert (eds.)