Triton X-100 Protein Lysis Buffer, autoclaving, urea etc..

Iraz Toprak Aydin via (by iraz.aydin from
Wed Jan 13 06:50:39 EST 2010

Hello everyone!


I will soon try to isolate proteins from melanocytes, I need to get both
Triton X-100 soluble and insoluble fractions. For this purpose I will make 

 this lysis  buffer;  50 mM Tris,  150 mM NaCl,  1% Triton X-100,  10 mM
EDTA, pH 7.2. And for the insoluble fraction I will use the same buffer with
8M urea. So my questions are:


1.      Can I autoclave the lysis buffer? Or should I filter it? (22um or

2.      For the lysis buffer with urea, can I also prepare this one before
and store it? Or shall I add the urea before use?

3.      Can I prepare a urea stock solution? Which concentration and storage
conditions would you recommend?



Thanks in advance!


Iraz Toprak Aydin


EPFL SV ISREC, Station 19

Batiment SV, SV 2540  

CH-1015 Lausanne



Tel: +41 21 693 07 36


e-mail:   <mailto:iraz.aydin from> iraz.aydin from


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