Two-Dimensional Electrophoresis and Identification by Mass Spectrometry.
Two-dimensional electrophoresis (2-DE) is a classical separation technique for complex protein mixtures based on a first-separation step by isoelectric point (pI) followed by SDS-PAGE. Due to its high resolving power (including protein isoforms) and the capability to display post-translational modifications, 2-DE has been the technique of choice in many studies focused in characterizing allergens. In this chapter, we describe a standard protocol which enables efficient protein separation and subsequent protein identification by tandem mass spectrometry (MALDI-MS/MS).
Centre for Cardiovascular Science, Queen's Medical Research Institute, University of Edinburgh, Edinburgh, UK.
Fernando
F
de la Cuesta
Laboratorio de Inmunoalergia y Proteomica, Departamento de Inmunologia, IIS-Fundacion Jimenez Diaz, Avda. Reyes Catolicos 2, 28040, Madrid, Spain. galvarez@fjd.es.
Gloria
G
Alvarez-Llamas
Department of Vascular Physiopathology, Hospital Nacional de Parapléjicos, Toledo, Spain.
Maria G
MG
Barderas
D000485:Allergens / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006801:Humans; D011499:Protein Processing, Post-Translational / Q000502:physiology; D011506:Proteins / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods
D000485:Allergens; D011506:Proteins
2-DE; Allergens; MALDI-TOF; Mass spectrometry; Two-dimensional electrophoresis