Hello!
I recently tested a new precast gradiented gel for my Western Blot experiments and I obtained wavy bands both for proteins and for molecular weight marker. I run in tris-glycine-sds running buffers as suggested by the company. You can find the pic in attachment.To be honest I observed similar bands pattern also in home made gels and this is the reason why I tried to switch to precast. Do you have any idea about the reason why I obtain such a bands?
Thank you
Elisa
Hi
i think you did not uses a homogeneous gel and also you should double check your electric flow during the gel was same.
Check electrodes ( undesirable side-reactions- oxidizing ) and power сables
Electrophoresis requires the maintenance of a direct current (DC) or steady electric field in an electrolyte.
there are some possibilities:
1. use fresh running buffer, mostly it occurs when running buffer is old.
2. Make sure you mix your resolving and stacking gels properly before adding them to the assembly.
3. Temperature may increase during electrophoresis .. try it at cold/room temperature.
4. check voltage and current during electrophoresis, you can reduce the voltage.
5. the most important thing could be PH and salt precipitation of the gel. check it correctly.
Thank you all. About the possibility it could be for the gel solutions its not the case because I used a precast gel... maybe as you said it could be the power supply or the electrodes because they are a little bit old...I will check them
In my opinion buffers may be a problem. I would propose to check correct pH and conc. of chemicals, salts etc.
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