title: Detection and semiquantitation of albumin forms in fresh human plasma separated on gradient polyacrylamide gel by means of electroblotting on agarose gel matrix.
abstract: Albumin in vitro contains several molecular forms, while in vivo it exists mainly as a monomer with a small fraction of a dimer. The aim of the present work was to detect and estimate albumin forms in fresh blood samples. The available analytical methods at present are inadequate for this purpose. An improved immunoblotting method was used where plasma was subjected to electrophoretic separation on 4-25% gradient polyacrylamide gels followed by immunoblotting on agarose gel containing anti-human albumin. The interference from the huge amount of the monomer in plasma was overcome by cutting the monomer region from the polyacrylamide gel before immunoblotting. After staining of the agarose gel, it revealed the presence of seven stained bands of albumin in addition to the monomer. These bands represent albumin aggregates and complexes of varying molecular masses (112-428 kDa). These albumin forms accounted for 0.7% of the total plasma albumin and their estimated level was 30.7 mg/dL. This study shows that the native albumin in blood has several molecular forms. It is concluded that albumin in healthy human subjects may form association complexes of varying molecular masses with other macromolecules in blood and these complexes are expected to be of physiological relevance.
authors.affiliation: Department of Chemical Sciences, University of Science and Technology, Irbed, Jordan.
authors.forename: R F
authors.initials: RF
authors.lastname: Atmeh
authors.forename: B
authors.initials: B
authors.lastname: Shabsoug
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D015151:Immunoblotting; D046911:Macromolecular Substances; D008970:Molecular Weight; D012685:Sepharose; D012709:Serum Albumin / Q000032:analysis*
chemical_list: D046911:Macromolecular Substances; D012709:Serum Albumin; D012685:Sepharose