Identification of human blood plasma and serum proteins in two-dimensional gels by use of protein A-Sepharose: application to alpha 1-microglobulin.
The relative position identification of some of the proteins in human blood plasma and serum after separation by two-dimensional electrophoresis (2-DE) in gel slabs have been studied. Antibodies specific to the protein to be identified were first immobilized on Protein A-Sepharose. Serum or plasma samples devoid of this protein were obtained following adsorption and subsequently the protein itself was eluted. The preparations were examined by 2-DE, silverstaining and blotting with parallel analysis of the purified proteins. Position identification in 2-DE gels of the spots of alpha 1-microglobulin (= protein HC) is shown as an example. The technique has general applicability not only for identification of the positions in 2-DE gels of serum and plasma proteins but also for proteins in other fluids, tissues and organs from humans and other species.
Vesterberg
O
O
National Institute of Occupational Health, Solna, Sweden.
Anundi
H
H
D000510:Alpha-Globulins / Q000032:analysis*; D015415:Biomarkers; D001798:Blood Proteins / Q000032:analysis*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006801:Humans; D011480:Protease Inhibitors / Q000032:analysis; D012685:Sepharose / Q000031:analogs & derivatives*; D016622:Silver Staining; D013205:Staphylococcal Protein A
D000510:Alpha-Globulins; D015415:Biomarkers; D001798:Blood Proteins; D011480:Protease Inhibitors; D012685:Sepharose; C042030:Staphylococcal protein A-sepharose; D013205:Staphylococcal Protein A; C016001:alpha-1-microglobulin