protein did not enter the gel on EMSA - (Sep/26/2005 )
I am doing EMSA. but I found there is strong signal in the well. How does the protein:DNA complex not enter the gel, I use 5% non-denature polyacrylamide gel.
-zzlm-
There can be several reasons, the most simple one is if your wells are not clean, do you wash it with a syringe? The second reason can be if your protein is forming oligomers or aggregates - is it a purified protein or extract? try lowering the concentration of the protein. Lastly use 4% gel to see if it helps.
-Cricked_Up-
Hi!
I've got the same problem with mines. My EMSA are carried out with 120mM final NaCl. Perhaps, the ionic strength is not well gauged.
Nicolas From Paris
-stormrider-