Percoll Gradients

[Denise Carpenter]

Hi,
    Can anyone with any experience in percoll gradients and their use in 
fractionation of subcellular compartments please help me with protocols 
which work!  I plan to use pancreatic, hepatic and T cell lines to 
primarily identify endosomes, lysosomes and plasma membranes.  The protocols 
I have leave me with a smudge on marker enzyme activity along the gradient, 
ie. beta galactosidase, beta glucoronidase, and LDH.  I would be most 
gratful for some advice on spin times and g force required to separate these 
compartments to enable accurate identification.

Thanks in advance for any suggestions.

Denise Carpenter
Dept of Pharmacology
University of Western Australia
e-mail: dcarpenter at receptor.pharm.uwa.edu.au