SDS gel not polymerize - (Apr/11/2005 )

hi fella,
I had no problem about SDS page gel before. This is a new lab i came. my resolving gel does not polymerize , i made fresh APS. only one thing i concerned that whether TEMED can go off. this lab kept TEMED room tep which was kept in my previous lab 4 degree.
What is the problem might be?
Thanks
cathy

Hi cathy

you shoul store TEMED in dark.We keep it at room temperature and it works. you should also degass your solution for 5 minutes minimum for good polymerization. Do you put n-butanol after you pour your running gel solution?if you don't you should do.That's all I know I hope I could help you:)

Hi cathy

you shoul store TEMED in dark.We keep it at room temperature and it works. you should also degass your solution for 5 minutes minimum for good polymerization. Do you put n-butanol after you pour your running gel solution?if you don't you should do.That's all I know I hope I could help you:)

hi
i had this problem in the past. I made fresh APS and problems gave up.
Temed is in a dark bottle in the fridge in my lab. But according to BURCU reply,your temed may be still ok
fred

Cathy,

TEMED can go off but since it can be stored at RT while most labs keep at 4C, it probably is not the problem.

2 things to think about: a) if the lab is cool (below RT), it will take longer to polymerise the gel
and APS powder can absorb water and therefore your stock may not really be the correct %.

My suggestion ... get new APS powder and make new 10% stock.

PS: if pouring two-stage gels, butanol may give a nice visual line when the gel has set, but even water is fine to use. All this layering does is prevents oxygen from reacting with the gel thereby enabling the gel to set properly.

Hope this helps,

Aussie.

Hi again:)

I think the problem is in the storage of TEMED:) we store it with aluminum foil and keep it in dark closet.it is approximately 23 degree.And also we prepare fresh APS and keep it with aluminum foil again untill we use it. Have a nice work

problem solved. the APS and TEMED are both ok actrully. there are two bottles of acrylamine. one is 40% acrylamine/bis and another one is 40% pure acrylamine. i get the wrong bottle. by the way, what is the principle of crosslink of bis and acry?

Thanks for you guys reply. make me feel lots people are back up me.