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

[Iraz Toprak Aydin]

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
45um)

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

Switzerland

 

Tel: +41 21 693 07 36

 

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