Protein binding to nitrocellulose, nylon and PVDF membranes in immunoassays and electroblotting.
A selection of different membranes commonly used to bind proteins in blotting and dot binding assays were investigated for a range of properties which would influence their performance. Large differences were observed in the membranes' ability to bind increasing amounts of protein, the effect of incubation times on the quantity of protein bound and the loss of proteins from the membranes following their incubation with different detergents or protein blocking agents. These differences could only partially explain the observed performance of the membranes when used as protein adsorbants in immunoassays and when different buffers were used for the electro-transfer of several different proteins to a range of membranes.
Tovey
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ER
Kolling Institute, Royal North Shore Hospital, St. Leonards, Australia.
Baldo
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BA
D000941:Antigens / Q000032:analysis; D003101:Collodion; D007118:Immunoassay / Q000379:methods*; D015151:Immunoblotting / Q000379:methods*; D007202:Indicators and Reagents; D007700:Kinetics; D008567:Membranes, Artificial; D009757:Nylons; D011145:Polyvinyls; D011485:Protein Binding; D011506:Proteins / Q000032:analysis*; D011863:Radioimmunoassay / Q000379:methods
D000941:Antigens; D007202:Indicators and Reagents; D008567:Membranes, Artificial; D009757:Nylons; D011145:Polyvinyls; D011506:Proteins; D003101:Collodion