immunochimica · of a fused hybrid (scale bar: 13.3 um); (e, f) hybridoma colomes maintained in...
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IMMUNOCHIMICA
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ANTIGENE ANTICORPORISPOSTA
IMMUNITARIA
RATTO CONIGLIO
A B
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METODI IMMUNOCHIMICI
ELISA (enzyme-linked immuno-assorbent assay)
Western blot
Radioimmunotest
Immunoistochimica
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IMMUNOBLOTTIG (WESTERN BLOTTING)IMMUNOBLOTTIG (WESTERN BLOTTING)
1.1. Separazione delle proteine (caricateSeparazione delle proteine (caricatenegativamente) sulla base del peso molecolare connegativamente) sulla base del peso molecolare congel elettroforesigel elettroforesi
2.2. Trasferimento delle proteine separate dal gel adTrasferimento delle proteine separate dal gel aduna membrana di supporto (elettroforetico)una membrana di supporto (elettroforetico)
3.3. ImmunoImmuno--rilevazionerilevazione delle proteine di interessedelle proteine di interesse
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ELETTROFORESIELETTROFORESI
Macromolecole cariche positivamente oMacromolecole cariche positivamente onegativamente (acidi nucleici e proteine) sononegativamente (acidi nucleici e proteine) sonoseparate in un campo elettrico.separate in un campo elettrico.La velocitLa velocitàà alla quale una molecola migraalla quale una molecola migra èè::
••Proporzionale alProporzionale al voltaggio applicatovoltaggio applicato e allae alla caricacaricaelettrica della molecolaelettrica della molecola
••Inversamente proporzionale alleInversamente proporzionale alle dimensionidimensionidella molecoladella molecola e allae alla viscositviscositàà della soluzionedella soluzione
Matrici: AGAROSIO, POLIACRILAMMIDEMatrici: AGAROSIO, POLIACRILAMMIDE
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WESTERN BLOT
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Western Blot in Farmacologia
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Radioimmunotest
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CURVA DI CALIBRAZIONE
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RADIOIMMUNOTEST
O + Ab CARBONE 1000
BK no Ab CARBONE 100
1 +Ab CARBONE 900
CURVA 5 +Ab CARBONE 800
STANDARD 10 +Ab CARBONE 600
CCK 20 +Ab CARBONE 400
50 +Ab CARBONE 200
125I CCK cpm
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CURVA DI CALIBRAZIONE