Detection of serum proteins by native polyacrylamide gel electrophoresis using Blue Sepharose CL-6B-containing stacking gels.
Analysis of serum proteins by native polyacrylamide gel electrophoresis is difficult because albumin is abundant in serum and interferes with the resolution of other proteins, especially alpha-antitrypsin which has mobility that is very similar to that of albumin. We present here a method in which serum proteins are separated by polyacrylamide gel electrophoresis using stacking gels containing Blue Sepharose CL-6B, which has a high affinity for albumin, lipoproteins, kinases, and pyridine-nucleotide-dependent oxidoreductases. During electrophoresis, proteins that bind to Blue Sepharose CL-6B stay in the stacking gel and do not migrate into the separating gel. As a consequence, certain proteins, including alpha(1)-antitrypsin, can be detected as clear bands. This method overcomes the requirement for fractionation of serum samples prior to electrophoresis to remove albumin and allows the simultaneous analysis of many samples.
Department of Clinical Biochemistry, Kyorin University School of Health Sciences, Miyashita, Hachioji, Tokyo, Japan. murachan@kyorin-u.ac.jp
Haruhiro
H
Muratsubaki
Kaoru
K
Satake
Yasuhisa
Y
Yamamoto
Keiichiro
K
Enomoto
D001798:Blood Proteins / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D015151:Immunoblotting; D009391:Nephelometry and Turbidimetry; D012685:Sepharose / Q000031:analogs & derivatives*
D001798:Blood Proteins; C017842:cibacron blue-sepharose; D012685:Sepharose