Hi there,
Actually, I am very new in cell cytotoxicity assay. I am planning to check the anti-virulence activity of a compound (i.e. whether this compound can inhibit the production of Stx/Verotoxin) against E. coli (STEC). I am planning to do culture of the E. coli (1×107 CFU) with and without different concentration of the compound. After overnight culture, I will collect the supernatent and prepare cell lysate. Then, I will use the supernatent and cell lysate to see the effect on vero cells in Microtitre plates. But, I am little confused-
1. How long I should culture the E. coli with/without the compound? Whether, I should culture with/without shaking?
2. How should I prepare the vero cells in 96 well Microtitre plates and what would be the dose of supernatent and cell lysate to see their effect?
Please suggest me a protocol that will be easier and best for this study.
- Badges
- Science topic