SDS Gel for 2D -DIGE - Problems having SDS Page to polymerize (Oct/07/2008 )

Hi peeps,

I've been casting SDS-PAGE with the following conc and components:

40% Bis/Acrylamide
1.5M Tris-HCL, pH 8.8
20% SDS
H2O
TEMED
10% APS

Have been able to get well-casted gel until yesterday, the gel turned out to be a of a gluey texture and soft (it does polymerized, however it seems only partial). I tried again today using the same recipe and vol. of each component and the same result again.

Was quite puzzling as nothing of this has happened before.
I'm suspecting that it's either the TEMED or APS that caused the prob, however....it does polymerize..just not totally.

Could anyone provide the storage duration of an opened bottle of TEMED in 4 degree fridge?
And any idea to the above situation?

Thanks alot.

SnowCasp

I would agree, sounds like a problem of APS or TEMED, prepare new APS and try again. I used my TEMED for years in 4°C although I suspect it is better not to store it there soo long Can't say how long it will be good, ask the supplier to be sure.

If your TEMED have gone very yellow, then you would better buy a new ones.

Thank you for the suggestion!!!


Hmm...turned out that the problem lies with the new bottle of acylamide that I'm using.

Tried casting with another bottle of acrylamide n it turned out well....
All those washing of the gel caster is a killer!!!