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Direct chemiluminescent imaging detection of human serum proteins in two‐dimensional polyacrylamide gel electrophoresis
Author(s) -
Tao Qian,
Wang Zhenzhen,
Zhao Heping,
Baeyens Willy R. G.,
Delanghe Joris R.,
Huang Lingyun,
Ouyang Jin,
He Dacheng,
Zhang Xianhua
Publication year - 2007
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200700078
Subject(s) - haptoglobin , chemiluminescence , hemopexin , polyacrylamide gel electrophoresis , blood proteins , chromatography , proteome , chemistry , acute phase protein , polyacrylamide , luminol , microbiology and biotechnology , two dimensional gel electrophoresis , protein detection , gel electrophoresis , proteomics , biochemistry , biology , immunology , gene , materials science , heme , inflammation , enzyme , nanotechnology
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.