HI,
I have transfected A549 cells with a plasmid of interest. The transfectants have been stably selected and clonally separated. I am doing a plate based fluorescence assay with the transfectants and unfortunately, with every wash, I am seeing a major cell loss. I have tried using pre-warm buffers and operating the the plate reader at 37 degrees too, but that doesn't seem to change much.
Thanks
Ruchi
Hi,
Have you tried to adhere the cells using Cell-Tak. See attached (no commercial interest). We have used this in one of the protocols for isolated ventricular mouse cardiomyocytes:
Article A method to fix and permeabilize isolated adult mouse cardio...
Good luck.
@Sathya Srinivasan
What type of plate are you using? Ruchi Jain If using optiplates, please check whether these are compatible with cell adhesions. Otherwise try coating the wells with poly-lysine or collagen first for better adhesion.
I concur with Dr. Nabodita Sinha Poly-D-lysine, Collagen, or Attachment factor coating will prevent the cells loss.
Thanks, I am using regular cell culture plates. But will try the coating and look into CellTalk also. Thanks
- Badges
- Science method