Electrophoretic protein transfer applied to immunoelectrophoresis.
A method for electrophoretic transfer of a crossed immunoelectrophoretic (CIE) pattern from an agarose gel onto a nitrocellulose sheet is described. The CIE pattern to be transferred was formed using Nonidet P-40 solubilized, bovine herpes virus-1 (BHV-1) induced antigens and bovine hyperimmune serum. An autoradiogram of a CIE pattern, formed using [35S]methionine-labeled NP-40 solubilized, BHV-1 induced antigens and bovine hyperimmune serum, was used as a control pattern. Both antigen and antibody were transferred to the nitrocellulose sheet. This was demonstrated by the ability of transferred antigen to react with specific antibody detected by the addition of 125I-labeled anti-species antibody and transferred antibody to react with both [35S]methionine-labeled antigen and 125I-labeled protein A. This method has application for determining the specificity of monospecific sera for individual arcs in CIE patterns.
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis* / Q000276:immunology; D000937:Antigen-Antibody Reactions; D000956:Antigens, Viral / Q000032:analysis* / Q000276:immunology; D001345:Autoradiography; D002417:Cattle; D006566:Herpesviridae Infections / Q000276:immunology; D007122:Immunoelectrophoresis / Q000379:methods*; D007123:Immunoelectrophoresis, Two-Dimensional / Q000379:methods*; D011817:Rabbits; D012016:Reference Values; D014764:Viral Proteins / Q000276:immunology*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D014764:Viral Proteins