We concentrated all our effort to carry out two electrophoreses in polyacrylamide gels (PAGE):
- SDS-PAGE (polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate; denaturing conditions): proteins from pomegranate samples were separated according to their size in 12% acrylamide gels and, after electrophoresis, gels were stained both with Coomassie Blue, and then with silver nitrate. See how beautiful look our gel.
- Native PAGE (polyacrylamide gel electrophoresis under non-denaturing conditions): proteins were separated according to their size and charge in 10% acrylamide gels. After electrophoresis, gels were stained for superoxide dismutase (SOD, EC 1.15.1.1) activity. As we can see in the following picture, some samples contained up to four activity bands, whereas some others had up to six. This is the first time that SOD activity is detected in pomegranate. We have been lucky: our results are pioneer.
Now, we are ready to prepare our reporting work, either as a poster for the PIIISA congress, or to be presented as an oral presentation. We have to figure out how to continue this project and have to discuss thoroughly about our results.
I have a question related with the polyacrylamide gels.
ReplyDeletefour bands and six bands of superoxide dismutase activity in the extracts are many or few? How many were expected? How many present other extracts, other plants, other tissues??
Nice Project!!!
Dr Francisco Martínez-Abarca
This is an interesting question. Overall, the common number of SOD isozymes in most plant species varies from 3 to 4: 3 is quite usual; 4 is perhaps something unusual. SIX is totally uncommon, and basically related to plants which undergo some stress. But these plants were grown under normal environmental conditions, if we consider normal conditions those at about 1000 m altitude.
DeleteThis will be one of our topics for the future research