problem by using percoll to purify primary cell - (Dec/02/2003 )
when i do cell isolation, i use 20%/50% percoll to purify cell.
when add the cell suspension to the top of percoll, after centrifuge, the cell seem to clot together, finally the middle layer contains very little cell i wanted, but contain many other contamination cell types.
does anyone work on percoll , i am very apprecaited if someone can give me some comments
Percoll centrifugation will give a good result based on the speed, and the percentage (density) of gradient. Some time shape of the tube also affects.
You are using two step percoll gradient. One is 50% and other is 20%. There may be a factor of dilution when you add cell suspension. You can take care of the exact density of the percoll gradients you use.
Avoid mixing while you load low density on top of 50 %
Dont know what type of cell you are interested in. Anyway this is the basic knowledge I have regarding percoll to share with you