Application of two-color infrared fluorescence for detection of protein phosphorylation / 南方医科大学学报
Journal of Southern Medical University
;
(12): 150-153, 2006.
Article
in Chinese
| WPRIM
| ID: wpr-234174
ABSTRACT
<p><b>OBJECTIVE</b>To assess the value of two-color infrared fluorescence imaging system in detecting protein phosphorylation in comparison with chemiluminescent detection.</p><p><b>METHOD</b>The lung tissue homogenate of mice treated with lipopolysaccharide (LPS) for different time lengths were prepared to separate the proteins by SDS-polyacrylamide gel electrophoresis followed by transfer of the proteins onto PVDF membranes. The membranes were incubated with the antibodies against total p42/44 MAPK/phospho-p42/44 MAPK, and then with goat anti-mouse or anti-rabbit secondary antibodies conjugated to Alexa Fluor 680, IRDye 800 or horseradish peroxidase. The blotted proteins were detected and quantified using Odyssey infrared imaging system or chemiluminescent detection.</p><p><b>RESULTS</b>LPS treatment rapidly induced p42/44 MAPK phosphorylation, which reached the peak level 1 h after the treatment and resumed the baseline level at 12 h. Consistent results were obtained by the two detection methods, but two-color infrared fluorescence imaging system showed better sensitivity in detecting the target protein, and was easy to manipulate with good efficiency and capable of analyzing two proteins simultaneously.</p><p><b>CONCLUSION</b>Two-color infrared fluorescence system is a powerful system for detecting phosphorylation of proteins.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pathology
/
Phosphorylation
/
Shock, Septic
/
Chemistry
/
Lipopolysaccharides
/
Blotting, Western
/
Reproducibility of Results
/
Mitogen-Activated Protein Kinase 1
/
Mitogen-Activated Protein Kinase 3
/
Fluorescence
Type of study:
Diagnostic study
Limits:
Animals
Language:
Chinese
Journal:
Journal of Southern Medical University
Year:
2006
Type:
Article
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