title: Improved two-dimensional gel electrophoresis representation of serum proteins by using ProtoClear.
abstract: ProtoClear is a proprietary technique for clearing albumin and immunoglobulin G (IgG) from human serum samples. Albumin constitutes 57-71% of total serum protein and IgG ranges from 8-26%. Removal of these two proteins alone clears approximately 75% of the total protein present in serum and allows the detection of the remaining proteins that are present in far lower concentrations. ProtoClear effectively removed >95% of human serum albumin (HSA) and >97% of human IgG as measured by an anti-HSA competitive immunoassay and a radial immunodiffusion assay, respectively. ProtoClear was far more specific at removing albumin and IgG than Cibracon Blue Dye chromatography (Cibracon Blue), the typically utilized alternative. Comparing two-dimensional (2-D) gels of serum cleared by either Cibracon Blue or by ProtoClear, it was apparent that Cibracon Blue removed a number of proteins in addition to albumin. Following removal of albumin and IgG from serum, we found a significant improvement in the resolution of polypeptide spots detected on two-dimensional gels.
authors.affiliation: Proteomix, Inc., San Diego, CA, USA. blollo@tns.net
authors.forename: B A
authors.initials: BA
authors.lastname: Lollo
authors.forename: S
authors.initials: S
authors.lastname: Harvey
authors.forename: J
authors.initials: J
authors.lastname: Liao
authors.forename: A C
authors.initials: AC
authors.lastname: Stevens
authors.forename: R
authors.initials: R
authors.lastname: Wagenknecht
authors.forename: R
authors.initials: R
authors.lastname: Sayen
authors.forename: J
authors.initials: J
authors.lastname: Whaley
authors.forename: F G
authors.initials: FG
authors.lastname: Sajjadi
mesh_terms: D000418:Albumins / Q000302:isolation & purification; D001798:Blood Proteins / Q000032:analysis*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D011933:Reagent Kits, Diagnostic
chemical_list: D000418:Albumins; D001798:Blood Proteins; D007074:Immunoglobulin G; D011933:Reagent Kits, Diagnostic