hi,
I have used the following buffer for a variety of cell-typs:
10 mM Hepes-KOH pH 7.2
0.25 M Sucrose
1mM EDTA
1mM MgOAc
this buffer wont lyse your cells immediately,
instead, you push your suspension several times through a 21Gneedle
and/or through a cell cracker (depending on the cell type).
Always follow lysis with Trypan Blue staining, so you can make sure, you
give them just as much mechanical stress as they need to brake.
To my experience, this is the most gentle way possible.
Good luck, Ina
--
Ina Hinners
ICRF
Secretory Pathways Laboratory
44 Lincolns Inn Fields
WC2A 3PX London
UK
email: I.Hinners at icrf.icnet.uk
