PROCESS FOR IDENTIFYING AMYLOID PRECURSOR PROTEIN IN PROTEIN COMPLEXES OF HIGH MOLECULAR WEIGHT

The invention relates to a process for identifying native amyloid precursor protein in protein complexes of high molecular weight, isolated from cellular membranes, meant to be applied in Alzheimer's disease pathology. According to the invention, the process consists in separating the cell memb...

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Hauptverfasser: CODRICI ELENA, DUDĂU MARIA, POPESCU IONELA DANIELA, ANGHELACHE LAURENŢIU, ENCIU ANA-MARIA, MIHAI SIMONA, TANASE CRISTIANA
Format: Patent
Sprache:eng ; rum
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Zusammenfassung:The invention relates to a process for identifying native amyloid precursor protein in protein complexes of high molecular weight, isolated from cellular membranes, meant to be applied in Alzheimer's disease pathology. According to the invention, the process consists in separating the cell membranes from the other cell components, extracting the protein complexes with 0.5...1% detergent Triton-X 100 in 50...75 mM of imidazole buffer, followed by the separation of protein complexes by electrophoresis in polyacrylamide as gel, for 2...4 h, at constant amperage of 10mA/gel on ice, in electrophoresis buffer with a pH 8...10 and the complex transfer on transfer membrane, for 20...22 h, at constant amperage of 100 mA, on ice, followed by protein identification with specific antibodies. Invenţia se referă la un procedeu de identificare al proteinei precursoare a amiloidului în stare nativă, în complexe proteice cu greutate moleculară mare, izolate din membrane celulare, cu aplicare în patologia bolii Alzheimer. Procedeul, conform invenţiei, constă în separarea membranelor celulare de restul componentelor celulare, extragerea complexelor proteice cu 0,5...1% detergent Triton-X 100 în 50...75 mM tampon imidazol, urmată de separarea complexelor proteice prin electroforeză în gel de poliacrilamidă timp de 2...4 h, la amperaj constant de 10 mA/gel pe gheaţă, în tampon de electroforeză cuH 8...10, transferul complexelor pe membrană de transfer timp de 20...22 h, la amperaj constant de 100 mA, pe gheaţă, urmată de identificarea proteinei cu anticorpi specifici.