Direct chemiluminescent imaging detection of human serum proteins in two-dimensional polyacrylamide gel electrophoresis.
A novel chemiluminescence (CL)-based imaging method capable of directly detecting proteins in polyacrylamide gels after electrophoresis is proposed. Human serum proteins are presently detected by a direct CL imaging method after native 2-D PAGE. As a consequence, some proteins, including haptoglobin (Hp), Hp precursor, hemopexin (Hpx) precursor, Ig alpha-1 chain C region, and Complement C3 precursor can be detected and identified by MS and MS/MS techniques. These proteins are all acute phase proteins, which have been defined as biomarkers for certain diseases. Moreover, serum proteins from healthy people and cirrhotic patients were analyzed. A decrease in Hp spots for cirrhotic patients could be confirmed. The CL imaging conditions were optimized, including the concentrations of H(2)O(2) and luminol. The process of CL detection of proteins is simple, and there is no need for specialized equipment. In comparison with the traditional CBB-R250 staining method, the detection sensitivity was improved and the detection period decreased about 70 times. Hence, this technique possesses potentials as a rapid, convenient, and inexpensive analytical technique for protein detection and for the diagnosis of diseases.
Tao
Qian
Q
College of Chemistry, Beijing Normal University, Beijing, PR China.
Wang
Zhenzhen
Z
Zhao
Heping
H
Baeyens
Willy R G
WR
Delanghe
Joris R
JR
Huang
Lingyun
L
Ouyang
Jin
J
He
Dacheng
D
Zhang
Xianhua
X
D000595:Amino Acid Sequence; D015415:Biomarkers / Q000032:analysis; D001798:Blood Proteins / Q000032:analysis*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D013194:Staining and Labeling
D015415:Biomarkers; D001798:Blood Proteins; D010455:Peptides