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Testing for cell surface forms of class II major histocompatibility complex antigens and I i by radioiodination, biotinylation, and membrane immunofluorescence
Author(s) -
Elliott William L.,
Sorli Christopher H.,
Reisert Patricia S.,
Humphreys Robert E.
Publication year - 1989
Publication title -
american journal of hematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.456
H-Index - 105
eISSN - 1096-8652
pISSN - 0361-8609
DOI - 10.1002/ajh.2830300103
Subject(s) - biotinylation , immunofluorescence , histocompatibility , antigen , histocompatibility testing , major histocompatibility complex , cell , chemistry , microbiology and biotechnology , immunology , medicine , biology , antibody , biochemistry , human leukocyte antigen
Antibodies to either I i or class II major histocompatibility complex (MHC) antigens did not recognize cell surface forms of I i in immunoprecipitates of cells that had been radioiodinated by the lactoperoxidase method, whereas they bound [ 35 S]methionine metabolically labeled molecules. N‐hydroxysuccinimidobiotin (NHS‐B) and biotin hydrazide (B‐H) were used to react more generally with cell surface proteins via amino groups and nitrene coupling, respectively. Each of these latter compounds labeled α and β chains of class II MHC antigens as seen in Western‐blotted, electrophoresed immunoprecipitates probed with 125 I‐labeled streptavidin but not I i or its associated forms. Although tyrosine residues might have been inaccessible to radioiodination in carbohydratederivatized forms of I i , the lack of I i biotinylation in these controled, sensitive studies was consistent with the view that I i forms were not surface expressed, with the possible exception of the chondroitin sulfate‐derivatized forms of I i (I i ‐CS).