Immunoblotting techniques with picogram sensitivity in cerebrospinal fluid protein detection.
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.
Nespolo
A
A
Department of Clinical Investigations, Istituto Neurologico Carlo Besta, Milano, Italy.
Bianchi
G
G
Salmaggi
A
A
Lazzaroni
M
M
Cerrato
D
D
Malesani Tajoli
L
L
D002556:Cerebrospinal Fluid Proteins / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D015151:Immunoblotting / Q000379:methods*; D007074:Immunoglobulin G / Q000134:cerebrospinal fluid*
D002556:Cerebrospinal Fluid Proteins; D007074:Immunoglobulin G