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Fig. 1. Analysis of MHC class II molecules in SaI/Ak cells. (A) Surface expression of I-Ak molecules on SaI, SaI/Ak and SaI/Ak tr cells was determined by FACS analysis. Binding of I-Ak mAb and the isotype control Ig (IgG2b) is shown. (B) Peptide-associated MHC class II {alpha}ß compact heterodimers (C{alpha}ß), high-molecular-weight (HMW) complexes and free ß chains were detected by western blotting on total cell lysates from SaI, SaI/Ak and SaI/Ak tr cells. Lysates were incubated in Laemmli sample buffer for 30 minutes at room temperature (nb, lanes 1, 2) or for 10 minutes at 95°C (b, lanes 3 and 5). Samples were migrated on a 10% SDS-PAGE gel before transfer to a PVDF membrane and immunoblotting with 10.2.16 mAb.





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