I treated my cells with Bafetinib for 2h, 4h and 8hrs. I harvested my cells at the specified timepoints by scrapping the adherent cells in the media containing Bafetinib, followed by 2 washes with PBS and then cell lysis.
I wasnt aware that I have to remove the media containing Bafetinib and replace it with PBS before scrapping my cells.
Will this have any effect on my cells and my western blots?
Thanks!
I think you are still ok since you did wash with PBS. It is more common to wash first but if cells are stressed and washing will remove them then scraping first and washing the pellet should be fine.
Agreed with Julie Ann Dougherty , we did have protocol that wash after the cells were been collected, the result showed no significant difference compared to the one that washed first in the same cell line and treatment.
This question lacks information, I think you should specify the type of cell you want to culture and which drug you are talking about.
Anyway, some drugs are likely to alter the pH of the medium to either acidic or basic. This obviously inhibit the growth of cell whose optimum pH is in contrast. Other drugs simply increase the osmotic pressure and the cell is subjected to becoming flaccid instead of successful growing.
The most important in the culture is to know the type of cell to be cultured and the conditions in which it grows best. If these are unknown, then generate hypothesis followed by control experiments and find out.
Thanks.
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology
- Virology
- Virus