Rapid blotting of IgG to nitrocellulose with minimal IgM contamination.
A rapid method is described for the non-electrophoretic transfer of IgG from 0.5 mm thick agarose gels to nitrocellulose. Since the agarose gels are attached to a solid support, the blot is unidirectional. However, 90% of the IgG is transferred in 10 min with no visible loss of resolution. In this procedure, less than 2% of the IgM antibodies are transferred from the gel to the nitrocellulose membrane. Therefore, this technique can be used in IgG spectrotype analysis or antigen-specific assays without the prior removal of IgM antibodies. Approximately 5-6 h are needed to run the gels, blot, and develop the protein pattern with double-antibody immunoperoxidase staining.
D003101:Collodion; D004587:Electrophoresis, Agar Gel; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis*; D007075:Immunoglobulin M / Q000032:analysis; D007163:Immunosorbent Techniques; D007526:Isoelectric Point
D007074:Immunoglobulin G; D007075:Immunoglobulin M; D003101:Collodion