We've had a look at your gel in the office at work, and among the various thoughts that came to mind were the following.
1) Too much protein present per lane.
2) Did you centrifuge your samples to separate solubilised proteins from insoluble material after the sonication step or is this raw lysate?
3) Is there detergent in your sample? The streaky nature of the left hand end of the bands is reminiscent of what you can see with NP40 or SDS crystals being left behind in the wells
4) DNA contamination (related to point 1 & 2 above)
Can you describe the exact buffer and protocol you have used to prepare the samples above so that we can have a better idea of the conditions? This should allow suggestions to be made of what parameters to try optimising.
Best of luck.
If it was overstained as a result of the staining process or inadequate de-staining the entire gel would feature heavy background - there are regions of this gel that are essentially clear / colourless (relative to the stained bit), suggesting that the problem is either the gel itself, or most likely what has been run on it.
If there is more information about the gel, the protein samples, and the staining method (Conventional Coomassie in Methanol / Acetic Acid / vs Colloidal Coomassie in acidified aqueous solution) then we might be able to suggest solutions to the issues in hand.
- Badges
- Science method