Measurement of peptide dissociation from MHC class II molecules.
This unit describes a method of measuring the kinetic stability of complexes formed between purified MHC class II molecules and antigenic N-terminal fluorescein-labeled peptides. An HPLC-SEC allows for the separation of fluoresceinated MHC class II:peptide complexes from excess free dissociating peptides, while the fluorescence detector allows for the quantification of the remaining complexes throughout the dissociation time-course.
Chaves
Francisco A
FA
University of Rochester, Rochester, New York.
Sant
Andrea J
AJ
University of Rochester, Rochester, New York.
D000818:Animals; D017951:Antigen Presentation; D001671:Biochemistry / Q000295:instrumentation / Q000379:methods*; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D019793:Fluorescein; D006207:Half-Life; D000949:Histocompatibility Antigens Class II / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D016056:Immunodominant Epitopes; D051379:Mice; D046912:Multiprotein Complexes / Q000276:immunology / Q000378:metabolism; D010446:Peptide Fragments / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D011485:Protein Binding
D000949:Histocompatibility Antigens Class II; D016056:Immunodominant Epitopes; D046912:Multiprotein Complexes; D010446:Peptide Fragments; D019793:Fluorescein