We are having trouble with loosing the top of some of our lanes on SDS gels. Any ideas why?
How interesting. It appears that the affected lanes actually ran normally because the bands lower down match up with the unaffected lanes. It is as if the top part of the gel isn't actually staining properly. Is this a common occurrence? It might be worth loading pre-stained markers (preferably rainbow markers) into the lane along with your sample in the center lanes. That way you will know that the gel ran properly and that it is a staining problem. I will keep thinking about it, though!
P.S. I hope you and Bob and the pup(s?) are doing well!
It would appear that something has prevented the gel from staining. Looking at the lower section of the gel, it is highly likely the proteins are present and simply unstained.
What type of stain are you using?
Did you put a sponge or a piece of paper to help with the destaining? Or you had something in the tray while staining in the top of those band?
Maybe the sponge was on top of where you don't see bands now, and this part has destained more...
Or the gel is not homogeneous in that part of the gel
Further information:
The samples were protease K treated, TCA precipitated, acetone washed and boiled. When the same samples were rerun on another gel, the gel was either fine (no disappearing bands) or a different place on the gel disappeared.
It is not a staining problem because the bands were radioactive and the missing area on the gel do not produce any labeled band either.
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