SDS-Polyacrylamide Electrophoresis and Western Blotting Applied to the Study of Asthma.
Western blotting is used to analyze proteins after being separated by electrophoresis and subsequently electro-transferred to a membrane. Once immobilized, a specific protein can be identified through its reaction with a labeled antibody or antigen. It is a methodology commonly used in biomedical research such as asthma studies, to assess the pathways of inflammatory mediators involved in the disease.Here, we describe an example of western blotting to determine the factors involved in asthma. In this chapter, the methodology of western blotting is reviewed, paying attention on potential problems and giving interesting recommendations.
Department of Clinical Genetics, University Hospital of Navarra, Pamplona, Spain. virgarciasol@yahoo.es.|Salamanca Institute for Biomedical Research (IBSAL), Salamanca, Spain. virgarciasol@yahoo.es.
Virginia
V
García-Solaesa
Departament of Molecular Biology, Centro de Análisis Genéticos (Citogen), Zaragoza, Spain.
Sara Ciria
SC
Abad
D001249:Asthma / Q000378:metabolism*; D015415:Biomarkers / Q000032:analysis; D015153:Blotting, Western / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans
D015415:Biomarkers
Antibody; Electrophoresis; Horseradish peroxidase; PVDF membranes; SDS-polyacrylamide; Semidry electrophoretic transfer; Western blotting