Improved two-dimensional gel electrophoresis representation of serum proteins by using ProtoClear.
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.
Lollo
B A
BA
Proteomix, Inc., San Diego, CA, USA. blollo@tns.net
Harvey
S
S
Liao
J
J
Stevens
A C
AC
Wagenknecht
R
R
Sayen
R
R
Whaley
J
J
Sajjadi
F G
FG
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
D000418:Albumins; D001798:Blood Proteins; D007074:Immunoglobulin G; D011933:Reagent Kits, Diagnostic