2-D gel densitometer for high-contrast and selective imaging of chlorophyll-containing protein complexes separated by non-denaturing polyacrylamide gel electrophoresis
Language English Country Netherlands Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
12088887
DOI
10.1016/s0165-022x(02)00029-5
PII: S0165022X02000295
Knihovny.cz E-resources
- MeSH
- Electrophoresis, Gel, Two-Dimensional methods MeSH
- Chlorophyll analysis MeSH
- Electrophoresis, Polyacrylamide Gel methods MeSH
- Photosynthetic Reaction Center Complex Proteins isolation & purification MeSH
- Hordeum growth & development MeSH
- Light-Harvesting Protein Complexes MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Chlorophyll MeSH
- Photosynthetic Reaction Center Complex Proteins MeSH
- Light-Harvesting Protein Complexes MeSH
In this work, we present a home-made two-dimensional (2-D) CCD imaging system for the monochromatic densitometry of plane gels and its application to the imaging and densitometry of chlorophyll (Chl)-containing proteins separated by non-denaturing polyacrylamide gel electrophoresis. The monochromatic imaging of separated green bands at the wavelengths corresponding to their absorption band increases their contrast. This allows a better visualization of the faint-green bands in the gel and using of samples with lower Chl content for the electrophoresis. By the comparison of 2-D densitograms of the same gel illuminated with 670 and 650 nm lights, that is, at the red absorption maximum of Chl a and b, respectively, we achieved a selective imaging of the complexes with different Chl a/b ratio. This approach was used to specify an unknown band that appeared in the gel of the sample prepared from the thylakoid membranes of preheated barley leaves.
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