Immunoblotting techniques with picogram sensitivity in cerebrospinal fluid protein detection | Zendy

Nespolo Angelo | Zendy; Bianchi Giorgio | Zendy; Salmaggi Andrea | Zendy; Lazzaroni Monica | Zendy; Cerrato Dario | Zendy; Tajoli Livia Malesani | Zendy

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Immunoblotting techniques with picogram sensitivity in cerebrospinal fluid protein detection

Author(s) -

Nespolo Angelo,

Bianchi Giorgio,

Salmaggi Andrea,

Lazzaroni Monica,

Cerrato Dario,

Tajoli Livia Malesani

Publication year - 1989

Publication title -

electrophoresis

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.666

H-Index - 158

eISSN - 1522-2683

pISSN - 0173-0835

DOI - 10.1002/elps.1150100109

Subject(s) - nitrocellulose , isoelectric focusing , agarose , chemistry , chromatography , electroblotting , biotinylation , latex fixation test , membrane , streptavidin , conjugate , biuret test , urea , biochemistry , antibody , biotin , biology , mathematical analysis , mathematics , immunology , enzyme

Agarose isoelectric focusing followed by blotting with nitrocellulose, nylon or polyvinylidene difluoride membranes, and immunochemical detection of cerebrospinal fluid IgG with various combinations of antisera, was evaluated. Polyvinylidene difluoride proved to be an easy‐to‐handle and reliable membrane for protein blotting. Among immunochemical visualization reactions, the most sensitive employed biotinylated goat anti‐human IgG followed by streptavidin colloidal gold conjugate and silver enhancement in 20% w/v urea, allowing a sensitivity of less then 1 picogram IgG/band.

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