title: Blotting from Immobilized pH Gradient Gels: Application to Total Cell Lysates.
abstract: Isoelectric focusing as used in the first dimension of two-dimensional gel electrophoresis separates protein isoforms such as those due to phosphorylation and acetylation. The immunoblotting method described here reveals this diversity by a one-dimensional separation. Using commercially available immobilized pH gradient plates or strips, the resolved proteins are transferred to PVDF membranes by diffusion and are probed with protein-specific antibody. The system is useful for monitoring changes of banding patterns and permits parallel processing of samples. Since the effect of posttranslational modifications on the isoelectric point can be predicted, inferring the number and extent of modifications is possible.
authors.affiliation: Institute of Pharmaceutical Sciences, ETH Zurich, HCI H430, Vladimir-Prelog-Weg 1-5/10, 8093, Zurich, Switzerland, harry.towbin@pharma.ethz.ch.
authors.forename: Harry
authors.initials: H
authors.lastname: Towbin
mesh_terms: D000818:Animals; D005782:Gels; D015151:Immunoblotting / Q000379:methods*; D007525:Isoelectric Focusing; D007962:Leukocytes / Q000166:cytology*; D008567:Membranes, Artificial; D011145:Polyvinyls / Q000737:chemistry; D051381:Rats; D005465:Fluorocarbon Polymers
chemical_list: D005782:Gels; D008567:Membranes, Artificial; D011145:Polyvinyls; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers