I'm using Percoll to remove dead cells from cultured lymphocytes, but I don't know which density gradients to use and which layers to collect. Can anyone give me some advice?
Depends if you are culturing human or mouse cells. Different densities are required. Mouse naive splenocytes we used density 1.119. After activation use density 1.077.
Human cells? We use lymphopaque for everything but I must say it doesn't work very well. You lose a lot of cells.
Agree with Dr Deobagkar on the layer to collect and the subsequent steps.
just density 1.077? someone told me that i have to put another layer of density 1.043 to remove debris, that is the cells between density 1.077 and density 1.043 should be collected. Is that right?