problem in my native gel - (Jul/15/2006 )

hi every one,
i got the bands for the gel which i ran at 14% . now iam try to elute out the lower molecular weight protein from the same sample for which i have to firest seperate the protein of interest by native gel and have to run the colum for the specific molecular weight . but i have a problem that is my 16% gel is cracking out when it is run and gets fully fragile if it is put in staining solution. why is that and what is the remedy for it. it would be appriciabl if i get a favourable response

shankar

-shanker-

Why are you trying to elute from a gel? Don't you have access to HPLC? Gel filtration or ion exchange would keep your protein native, and should allow you to separate it from other protein.

-swanny-

You can reduce bis-acrylamide concentration a bit.

-genehunter-1-

QUOTE (swanny @ Jul 17 2006, 07:55 AM)

Why are you trying to elute from a gel? Don't you have access to HPLC? Gel filtration or ion exchange would keep your protein native, and should allow you to separate it from other protein.

i actually dont have the facilities for hplc and other things what you said in my lab. thats the prob for me now. i can ony do column

-shanker-

QUOTE (swanny @ Jul 17 2006, 07:55 AM)

Why are you trying to elute from a gel? Don't you have access to HPLC? Gel filtration or ion exchange would keep your protein native, and should allow you to separate it from other protein.

i dont have facilites for that in my lab

-shanker-