Imunokemijsko določanje invariantne verige poglavitnega histokompatibilnostnega kompleksa (MHC) II. razreda ter kolokalizacija s katepsinoma L in S v bioloških vzorcih : magistrsko delo = Immunochemical determination of invariant chain associated with major histocompatibility complex (MHC) class II and its co-localization with cathepsins L and S in biological samples : master thesis
Zavašnik-Bergant, Tina
The aim of the present study was to prepare specific mouse monoclonal antibodies directed against MHC class II-associated p41 invariant chain. BALBžc mice were immunized with human recombinant ...invariant chain, comprising only the luminal domain (isoform p26). For the preparation of hybridomas, mouse splenocytes and myeloma cells (NS 1) were fused. Hybridoma cells were screened for antibody production using ELISA and immunoblotting. Selected hybridomas were cloned twice by limiting dilution and monoclonality was confirmed by isoelectric focusing and subclass determination. Three cell linesproduced specific antibodies which reacted with p41 fragment. Monoclonal antibodies were purified by affinity chromatography on Protein A-Sepharose. Antibodies from selected cell lines showed little or no cross-reactivity against cathepsin L and cathepsin S. Antibodies from clone 2C 12 were used forimmunohistochemical analysis of MHC class II-associated invariant chain in lymph-node tissue together with antibodies against cathepsin L and cathepsin S. In lymph-node tissue specific localization of invariant chain, cathepsin S and cathepsin L was observed. In certain cells co-localization of invariant chain together with cathepsin S, but not with cathepsin L, was confirmed usingconfocal fluorescence microscopy. Cathepsin S was detected in subcapsular(cortical), medullary and trabecular sinuses of lymph node togetherwith tissue histiocytes (connective tissue macrophages). Localization of invariant chain and cathepsin S in lymph-node cells was confirmed by electron microscopy. In transformed Hodgkin cells and Stenberg-Reed giant cells of lymph-node tissue affected by Hodgkin's disease strong immunofluorescence staining for invariant chain and cathepsin S was observed.
Vrsta gradiva - magistrsko delo
Založništvo in izdelava - Ljubljana : [V. Zavašnik- Bergant], 2000
magistrske naloge ZAVAŠNIK-Bergant V. Imunokemijsko IN: 020001296
prosto - na dom, čas izposoje: 1 mes.
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