Chemiluminescent-based methods to detect subpicomole levels of c-type cytochromes.
A variety of luminol-based substrates and either an autoradiographic film or a charge-coupled device (CCD) imaging system were used for chemiluminescence detection of c-type cytochromes. The Pierce Femto peroxidase substrate was at least 10 times more sensitive when using film than the highly cited 3,3('),5,5(')-tetramethylbenzidine (benzidine derivative) staining method and 50 times more sensitive when using a CCD imaging system. Film or CCD imaging result in highly sensitive and quantitative signals. The quantitative detection of c-type cytochromes from single colonies or from less than 1ml of a bacterial culture is possible.
Feissner
Robert
R
Department of Biology, Washington University, St. Louis, MO 63130, USA.
Xiang
Youbin
Y
Kranz
Robert G
RG
D001426:Bacterial Proteins / Q000032:analysis; D001560:Benzidines / Q000737:chemistry; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002863:Chromogenic Compounds / Q000737:chemistry; D045304:Cytochromes c / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D006418:Heme / Q000737:chemistry; D006571:Heterocyclic Compounds / Q000737:chemistry; D008163:Luminescent Measurements; D010544:Peroxidases / Q000737:chemistry; D016354:Rhodobacter capsulatus / Q000254:growth & development / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods
D001426:Bacterial Proteins; D001560:Benzidines; D002863:Chromogenic Compounds; D006571:Heterocyclic Compounds; C021758:3,3',5,5'-tetramethylbenzidine; D006418:Heme; D045304:Cytochromes c; D010544:Peroxidases