title: Post-stained Western blotting, a useful approach in immunoproteomic studies.
abstract: The precise localisation of immunogenic proteins on stained two-dimensional electrophoresis (2DE) gels is occasionally difficult, contributing to the erroneous identification of unrelated non-immunogenic proteins, which is expensive and time consuming. This inconvenience can be solved by performing immunoblotting using previously stained polyacrylamide gels. This approach was proposed nearly 20 years ago but is now almost forgotten. We have evaluated the suitability of this approach to identify immunogenic proteins from Lactococcus garvieae. Some of the immunogenic proteins identified in L. garvieae, such as Gls24, have been considered important as immunotarget in different bacterial species. Post-staining western blotting facilitated the correct selection of immunogenic proteins of interest in 2D gels before their identification.
authors.affiliation: Department of Animal Health, Faculty of Veterinary Science, Complutense University, 28040 Madrid, Spain.
authors.forename: Mercedes
authors.initials: M
authors.lastname: Reguera-Brito
authors.affiliation: Department of Animal Health, Faculty of Veterinary Science, Complutense University, 28040 Madrid, Spain; Animal Health Surveillance Center (VISAVET), Complutense University, 28040 Madrid, Spain.
authors.forename: José F
authors.initials: JF
authors.lastname: Fernández-Garayzábal
authors.affiliation: Department of Animal Health, Faculty of Veterinary Science, Complutense University, 28040 Madrid, Spain.
authors.forename: M Mar
authors.initials: MM
authors.lastname: Blanco
authors.affiliation: Department of Animal Health, Faculty of Veterinary Science, Complutense University, 28040 Madrid, Spain.
authors.forename: Mónica
authors.initials: M
authors.lastname: Aguado-Urda
authors.affiliation: Department of Animal Health, Faculty of Veterinary Science, Complutense University, 28040 Madrid, Spain. Electronic address: gibelloa@vet.ucm.es.
authors.forename: Alicia
authors.initials: A
authors.lastname: Gibello
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis* / Q000276:immunology; D015153:Blotting, Western / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional; D006360:Heat-Shock Proteins / Q000032:analysis* / Q000276:immunology; D017045:Lactococcus / Q000737:chemistry* / Q000276:immunology; D058977:Molecular Sequence Annotation; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013194:Staining and Labeling
chemical_list: D001426:Bacterial Proteins; D006360:Heat-Shock Proteins
keywords: Detection; Immunogenic protein; Post-staining; Western blot