separation of loading buffer - (Nov/18/2008 )
hi!
could u help me?
when i run my sample in SDS gel,
my enzyme and the loading buffer will separate,
so when it passes the gel,
it seems that my sample is in the upper part of the loading buffer.
I run in 80V only.
and anyone know how to increase the intensity of band?
i try to incubate and prolong the time of staining, but it cant significantly sharpen the band.
thanks in advance
-vivale-
QUOTE (vivale @ Nov 18 2008, 12:44 AM)
hi!
could u help me?
when i run my sample in SDS gel,
my enzyme and the loading buffer will separate,
so when it passes the gel,
it seems that my sample is in the upper part of the loading buffer.
I run in 80V only.
and anyone know how to increase the intensity of band?
i try to incubate and prolong the time of staining, but it cant significantly sharpen the band.
thanks in advance
could u help me?
when i run my sample in SDS gel,
my enzyme and the loading buffer will separate,
so when it passes the gel,
it seems that my sample is in the upper part of the loading buffer.
I run in 80V only.
and anyone know how to increase the intensity of band?
i try to incubate and prolong the time of staining, but it cant significantly sharpen the band.
thanks in advance
what do you mean exactly that your enzyme separates? you mean the loading dye runs faster? it is always like that.
pH is very important in the quality of your results and bands that you see on your gel. check the pH of all your buffers.
also, I don't think treatment and boiling for a longer time in SDS-PAGE buffer would sharpen your bands.
-Curtis-