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Dot blot positivo, Wb no!

Dot blot positivo, Wb no! - Forum Biologia Molecolare

Dot blot positivo, Wb no! - Forum Biologia Molecolare. Speak Italian Molecular Biology Research Forum.


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Old 07-03-2009, 08:36 AM
Pipette Filler
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Default Dot blot positivo, Wb no!



Salve a tutti!

da tempo cerco su omogenati di cervello un'enzima (P450C17, aromatasi per la sintesi di DHEA) mediante WB ma senza alcun risultato. Il protocollo che uso validato (ho lavorato con altre proteine) e mi sento di escludere problemi tecnici sostanziali (anche perh fino al trasferimento, semi-dry, come ho sempre verificato con il Ponceau tutto ok).
La cosa che mi lascia perplesso aver comunque rilevato un segnale mediante Dot-Blot usando stesso anticorpo, alle stesse concentrazioni sullo stesso omogenato. Sto intanto verificando P450C17 con IF su brain slices nelle stesse regioni ma nel frattempo, per quanto riuarda il WB, non so davvero che pesci prendere...magari far un'immunoprecipitato prima del prossimo WB...
Vi chiedo quindi se avete qualche idea e/o suggerimento.

Grazie!
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blot , dot , dot blot positive , immunoprecipitation , p450c17 , positivo , wb negative


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