I have influenza virus produced in eggs and then purified by ultracentrifugation on a sucrose gradient after protein determination to know the amount of virus to put in the protein gel I run the gel and then stain either with coomassie or silver staining.
We are getting very difficult to read gels. Any advice on how to get a cleaner and nicer run for analysis without doing western?
Thank you for your help.
Try the same protocol which you use for SDS PAGE of a recombinant protein. Since you are using the ultracentrifuged allantoic fluid, it is likely that you will get the other protein bands corresponding to the proteins present in allantoic fluid.
You can confirm those bands by western blotting.
If you wish to get only virus specific bands in SDS-PAGE, try density gradient centrifugation to get purified virus.
Secondly keep your SDS-PAGE loading volume towards higher side (10-15ul of 8ml ultracentrifuged virus- AF titre >26 gives very good band intensity).
Good luck
Influenza viruses have a tendency to to get aggregated in solution. In the past we have sonicated the samples before centrifugation to reduce nonspecific proteins binding to the viruses
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