gel shift assay help- big smear lane - (May/18/2006 )
Dear all
I am working on gel shift. unfortunately I saw the whole lane big blank, could not see any binding. I am sure that should have some binding. I could not figure what is wrong with it.
gel? loading? or anything? anyone give me some advise.
Thanks millons
-cathy-
QUOTE (cathy @ May 18 2006, 03:38 AM)
Dear all
I am working on gel shift. unfortunately I saw the whole lane big blank, could not see any binding. I am sure that should have some binding. I could not figure what is wrong with it.
gel? loading? or anything? anyone give me some advise.
Thanks millons
I am working on gel shift. unfortunately I saw the whole lane big blank, could not see any binding. I am sure that should have some binding. I could not figure what is wrong with it.
gel? loading? or anything? anyone give me some advise.
Thanks millons
Did you see unbound probe at the bottom of lane? If not then your probe is degraded. The binding buffer (salt concentration, pH, DTT concentration) makes a big difference also.
Describe what you have done in more detail.
JP
-JP2877-
could also be quality of the extract...would you please post a picture for us?
-aimikins-