Recipe for 8% SDS PAGE - (Jan/16/2007 )

Hi everyone,

as I need a recipe for a 8% SDS-gel for Tricine SDS-PAGE I'm wondering if you could help me.
First I tried to recalculate from values of my 10% and 12% gels but that did not really work out at least for the water part.
Water and Acrylamid are the only factors that can be changed. So in that case of the following recipe, how many ml of
Acrylamid and Water I have to use for a 8% gel?

My recipes: 10% --- 12%

Acrylamid: 11,3 ml --- 13,6 ml
ddH2O: 11,3 ml --- 8,9 ml
gel buffer: 11,3 ml
APS (20%): 262,5 µl
Temed: 9 µl

Thanks for your replies!!!

Hi,

In my lab I used to prepare 30mL of separating mix as following (for 8%):
* 8mL Acrylamide 30% (29:1)
* 7.5mL Separating buffer 4X
* 14.5mL ddH2O
200uL APS 10% + 20 uL TEMED

Soooo for your volume (~33.9mL total volume) it will be something like this:
* 9.04mL Acrylamide
* 8.475mL Separating buffer 4X
* 16.385mL ddH2O
Then your TEMED and APS as usual