title: Efficient transfer of proteins from acetic acid-urea and isoelectric-focusing gels to nitrocellulose membrane filters with retention of protein antigenicity.
abstract: A method which facilitates the rapid and quantitative electrophoretic transfer of proteins from gels not containing sodium dodecyl sulfate (SDS) to nitrocellulose membranes is described. The equilibration of non-SDS-polyacrylamide gel electrophoretic gels in a buffer containing SDS confers a net negative charge to the proteins present, presumably as a result of the formation of SDS-protein complexes. Proteins from gels equilibrated in the SDS buffer and then electroblotted in a Tris-glycine buffer at pH 8.3 are transferred with much greater efficiency than are proteins from untreated gels. The method has been shown to significantly enhance the electrophoretic transfer of polyoma viral proteins resolved in either acetic acid-urea or isoelectric-focusing gels to nitrocellulose membranes, and it is suggested that the method should have universal applicability to all gel electrophoresis systems currently employed. The proteins from isoelectric-focusing gels treated with SDS and transferred to nitrocellulose membranes were found to retain antigenicity to antisera prepared against either denatured or native viral proteins.
mesh_terms: D000085:Acetates; D000941:Antigens; D003101:Collodion; D007525:Isoelectric Focusing / Q000379:methods; D008567:Membranes, Artificial; D011506:Proteins / Q000276:immunology / Q000302:isolation & purification*; D014462:Ultrafiltration; D014508:Urea
chemical_list: D000085:Acetates; D000941:Antigens; D008567:Membranes, Artificial; D011506:Proteins; D014508:Urea; D003101:Collodion