Hi Lina,

In our lab, the treatments are done on adherent cells before detaching them from the cultureware. Conjugated antibodies or dyes are added to the cell suspension before flow. Hope this helps.

Thanks alot sweelin. I understand that the cells get the treatment when they are relaxed in their normal condition.

I am just afraid of ligand to be washed out from the cells during washing steps.

As the ligand will affect the localization of the rexeptos.

What is your opinin?

Treatment should be before detachment because:

1. Addition of EDTA/Trypsin may affect the cell membrane thereby affecting the binding.

2. Anything that is bound on the receptor should not be easily washed away. The washing steps are essential to remove anything that has bound non-specifically. So whatever you are washing away is not what you want to acquire.

Thank you Darellynn

Darellynn's answer is to the point! You could also consider using a more gentle detachment solution like Accutase instead of trypsin which will not damage the surface receptors