| Stained protein on gel displays edge effects within gel lanes or band smearing. |
This could be a result of several problems. Adding 1 × sample buffer to any unused wells may often solve this problem. The problem may also stem from additives in the sample buffer, so one should minimize or decrease salts, detergents, and solvents during sample preparation and in sample buffers. |
| Sample is loaded onto gel, but sample floats out of the well. |
Add 10% glycerol to make sample denser than the surrounding buffer. |
| Gel does not destain quickly prior to cutting. |
The addition of one or two crumpled KimWipe tissue will bind residual Coomassie dye, and accelerate the destaining process. |
| Gel slice remains blue following several washes. |
Note that more frequent changes of Destaining Buffer may be necessary to sufficiently destain intensely-stained gel pieces. If gel pieces become dehydrated (white and rigid), destaining becomes less efficient. Alternating Destaining Buffer with a five-minute wash in organic solvent-free Digestion Buffer (100 mM EPPS pH 8.5) may aid the destaining process. |
| Following digestion, gel slices are dry. |
Ensure that the gel pieces are covered with sufficient buffer to account for evaporation so that the gels do not dry overnight. |
