problem of gelatin zymography testing mmp2/9 - (Mar/02/2006 )

Hi there

I followed all the procedure of chemicon protocol but i can never get "clear bands at blue backgroud". what i have is some faint bands. Help me out of here if you have any suggestion.
I used own made 8%SDS gel and 0.5mg/ml (or 1mg/ml) geltain. And I tried different incubation time and concentration of stain solution.


Thanks first

Hi there

I followed all the procedure of chemicon protocol but i can never get "clear bands at blue backgroud". what i have is some faint bands. Help me out of here if you have any suggestion.
I used own made 8%SDS gel and 0.5mg/ml (or 1mg/ml) geltain. And I tried different incubation time and concentration of stain solution.


Thanks first

thanks you, Veteran

I think the protein level is pretty high, 100ug/lane. And I got the clear band yesterday, but the size is not what i am expected, there are two close band around 130, not 92 and 75 which is mmp9 and mmp2. confused. are they dimmer or combined with other protein?

waiting for more suggestions