Ratio between cells and lysis buffer - (Sep/03/2008 )
Hi everyone
I am going to prepare a cell lysate from a cell line, but I have no idea on how much lysis buffer I should use to lysis the cells. Is there any ratio between no. of cells and the lysis buffer? Here is a recipe for my lysis buffer:
20mM Tris-HCl, pH7.5
150mM NaCl
1mM EDTA
1% Triton X-100
5mM iodoacetic acid
1mM PMSF
-Durandal-
QUOTE (Durandal @ Sep 3 2008, 03:51 AM)
Hi everyone
I am going to prepare a cell lysate from a cell line, but I have no idea on how much lysis buffer I should use to lysis the cells. Is there any ratio between no. of cells and the lysis buffer? Here is a recipe for my lysis buffer:
20mM Tris-HCl, pH7.5
150mM NaCl
1mM EDTA
1% Triton X-100
5mM iodoacetic acid
1mM PMSF
I am going to prepare a cell lysate from a cell line, but I have no idea on how much lysis buffer I should use to lysis the cells. Is there any ratio between no. of cells and the lysis buffer? Here is a recipe for my lysis buffer:
20mM Tris-HCl, pH7.5
150mM NaCl
1mM EDTA
1% Triton X-100
5mM iodoacetic acid
1mM PMSF
the more lysis buffer you use the more diluted are your proteins; if you use the 1x fold lysis buffer you have to use at least 5- to 10-fold volume of the volume of your cells...
to get concentrated lysates prepare 3- or more fold concentrated lysis buffer to use less lysis buffer
-The Bearer-