title: SDS-PAGE to Immunoblot in One Hour.
abstract: An ultra-rapid method for electrophoresing proteins on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transfer of proteins to nitrocellulose membranes, and immunoblotting is described here. Electrophoresis of the autoantigens La and Ro60, as well as molecular weight standards on a 4-20 % gradient gel, was performed in about 10 min using heated (70-75 °C) normal Laemmli running buffer. Electrophoretic transfer of these proteins was achieved in 7 min using a semidry transfer method. Finally, immunoblotting of La and Ro60 was carried out in 30 min. Thus, the entire process of electrophoresis, electrotransfer, and immunoblotting could be carried out in 1 h.
authors.affiliation: Arthritis and Clinical Immunology Program, Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK, 73104, USA, biji-kurien@omrf.org.
authors.forename: Biji T
authors.initials: BT
authors.lastname: Kurien
authors.forename: Debashish
authors.initials: D
authors.lastname: Danda
authors.forename: Michael P
authors.initials: MP
authors.lastname: Bachmann
authors.forename: R Hal
authors.initials: RH
authors.lastname: Scofield
mesh_terms: D003101:Collodion / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D008567:Membranes, Artificial; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification; D013997:Time Factors
chemical_list: D008567:Membranes, Artificial; D011506:Proteins; D003101:Collodion