Author: Kammer, Andreas R.; van der Burg, Sjoerd H.; Grabscheid, Benno; Hunziker, Isabelle P.; Kwappenberg, Kitty M.C.; Reichen, Jürg; Melief, Cornelis J.M.; Cerny, Andreas
Title: Molecular Mimicry of Human Cytochrome P450 by Hepatitis C Virus at the Level of Cytotoxic T Cell Recognition Document date: 1999_7_19
ID: 1vabzhs5_11
Snippet: Competition-based MHC Class I Peptide Binding Assay. Peptides were tested for their binding affinity using the previously described peptide binding assay (26) . In brief, cells were stripped (see above) and resuspended at 7 ϫ 10 5 cells/ml in IMDM plus 1.5 g/ml ⤠2 -microglobulin. A fluorescein (FL)-labeled reference peptide (FLPSDC(FL) FPSV), 25 l (end concentration, 150 nM), was incubated with 25 l of competitor peptide (different end concen.....
Document: Competition-based MHC Class I Peptide Binding Assay. Peptides were tested for their binding affinity using the previously described peptide binding assay (26) . In brief, cells were stripped (see above) and resuspended at 7 ϫ 10 5 cells/ml in IMDM plus 1.5 g/ml ⤠2 -microglobulin. A fluorescein (FL)-labeled reference peptide (FLPSDC(FL) FPSV), 25 l (end concentration, 150 nM), was incubated with 25 l of competitor peptide (different end concentrations) in a 96-well V-bottomed plate. 100 l of mild acid-treated JY cells was added to these wells. The mixture was incubated for 24 h at 4 Њ C, washed twice with PBS containing 1% BSA (PBA1%), resuspended in PBA1% containing 0.5% paraformaldehyde, and analyzed by FACScan ® (Becton Dickinson).
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