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Improved two‐dimensional gel electrophoresis representation of serum proteins by using ProtoClear TM
Author(s) -
Lollo Bridget A.,
Harvey Sheryl,
Liao Jane,
Stevens Anthony C.,
Wagenknecht Raymond,
Sayen Rick,
Whaley Justine,
Sajjadi Fereydoun G.
Publication year - 1999
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(19990101)20:4/5<854::aid-elps854>3.0.co;2-6
Subject(s) - radial immunodiffusion , albumin , chromatography , human serum albumin , chemistry , blood proteins , clearance , immunoassay , serum albumin , electrophoresis , immunoglobulin g , antibody , microbiology and biotechnology , biochemistry , biology , immunology , medicine , urology
ProtoClear TM 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 ˜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.