Catalase and superoxide dismutase double staining zymogram technique for Deinococcus and Kocuria species exposed to multiple stresses.
Superoxide dismutase (SOD) and catalase expression is associated with oxidative stress. Existing techniques for the individual staining of SOD and catalase have been described in the past. The objective of this study was to achieve a simple and rapid technique for the double staining of bacterial SOD and catalase on the same polyacrylamide gel. SOD detection was carried out using nitro-blue tetrazolium (NBT) dye reduction followed by ferricyanide precipitation for negative staining of the catalase enzyme on the same gel. The staining procedure resulted in pale blue SOD bands while catalase appeared as yellow bands against a greenish blue background on the same gel. This technique was used to detect changes in the polymorphic forms of these enzymes in Deinococcus radiodurans R1 and Kocuria sp. C2 subjected to stresses like UV and gamma radiation and desiccation.
Shukla
Manish R
MR
Department of Biological Sciences, Louisiana State University and A &M College, Baton Rouge, LA, USA.
Yadav
Radhika
R
Desai
Anjana
A
D002374:Catalase / Q000378:metabolism*; D034301:Deinococcus / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005292:Ferricyanides / Q000378:metabolism; D008835:Micrococcaceae / Q000201:enzymology*; D009580:Nitroblue Tetrazolium; D018384:Oxidative Stress; D020033:Protein Isoforms / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*; D013482:Superoxide Dismutase / Q000378:metabolism*
D005292:Ferricyanides; D020033:Protein Isoforms; D009580:Nitroblue Tetrazolium; D002374:Catalase; D013482:Superoxide Dismutase