Lysis on microtiter plate - (Oct/07/2001 )
I wants to know if it is possible to lyse cells growing directly in a 96 well plate for ELISA ?
The cells will be dying due to the treatment, so washing should be avoided. Can I disregard serum proteins, and will the cellular proteins coat the surface in presence of the lysis buffer ?
Thank you for your help
Here's a suggestion from a colleague:Reagents such as Triton will lyse the cells also in the 96-well format and in the presence of medium or serum. If you have serum in the medium, the surface will be already coated by serum proteins. So if dilution of cellular proteins and/or the presence of medium/FCS is not a concern, direct lysis is a good solution.Good luck
If your cells are adherent, then you are in better luck. If they are adherent you can allow the cells to grow directly on the plate, or I would coat the plate with Fibrinogen (or other adhesion molecules) to the plate O/N then allow the cells to bind to the Fibrinogen for a couple of hours. This allows you to wash your cells, just be VERY careful, I suggest aspirating with a very fine tip versus dumping. Then when you can add your antibodies for the ELISA. In your developing solution (be it for HRP or AP labelled Ab's) add a detergent (ie Triton X-100 or NP-40) to lyse the cells on the plate.