Measurement of cytochrome f in thylakoids of higher plants using quantitative gel electrophoresis.
A quantitative method was developed to estimate the concentration of cytochrome (cyt) f in isolated thylakoids, using sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by staining with a heme-specific reagent containing 3,3',5,5'-tetramethylbenzidine and hydrogen peroxide. This densitometric technique was at least as sensitive as difference spectroscopy. Analysis of thylakoid preparations by densitometry of stained bands using cyt c as standard gave molar ratios of cyt/chlorophyll which were identical to ratios obtained by difference spectroscopy. Densitometric assays demonstrated that the molar ratio of cyt f/chlorophyll decreased during leaf aging in seven higher plants; however, there was a marked difference in the rate at which cyt f was lost from the leaves of different species.
D001560:Benzidines; D002734:Chlorophyll / Q000032:analysis; D002736:Chloroplasts / Q000032:analysis*; D003580:Cytochromes / Q000032:analysis*; D045348:Cytochromes f; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D007887:Fabaceae; D001467:Hordeum; D010946:Plants, Medicinal; D013025:Glycine max; D013194:Staining and Labeling; D013997:Time Factors; D014908:Triticum
D001560:Benzidines; D003580:Cytochromes; D002734:Chlorophyll; C021758:3,3',5,5'-tetramethylbenzidine; D045348:Cytochromes f