lysis buffer / protein concentration

Joel Baguet Joel.Baguet at ens-lyon.fr
Mon Jul 21 09:11:28 EST 1997


Try to dialyze your samples on a mini column (eppendorff tube 1,5ml 
with a hole) on an other eppendorff tube with Sepharose CL-6B
-> 30 mcl of glass siliconized beads (Sigma G 1277) with a blue 
eppendorff tip
-> 600 mcl of sephadex G50 or G75 (2/3) in Tris 20 mM pH 7.5 or 8.0 
-> centrifuge 4 min at 3000rpm
-> change the bottom tube for a low binding tube 1.5 ml
-> put 50 mcl of cellular extract in lysis buffer on the top of the 
column
-> centrifuge 4 min at 3000 rpm
-> add 50 mcl buffer if the 50 mcl sample is not entirely passed 
through the column

it works well!!!

or try  an other system using 1 ml syringue with glass-wool at the 
bottom, 1 ml of sephadex G50 or G75 (1/2) in Tris 20 mM pH 7.5 or 8.0
and centrifuge 2000rpm I believe instead of 3000 rpm, the syringeu is 
on a little 0.7ml tube. All the system is in a 15 ml tube.



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