In our laboratory we have a problem with SDS Page technique, we have tried different sample concentration, the buffer that we use isn't expired and the pad that we put in the chamber is MOPS 1x, the voltage selection is 200. In every sample there is 20 ug of proteins the first time, and 30 ug the second time; the result is always the same. I attach the photo of the running gel. The chamber brand is Invitrogen Novex Mini-Cell.
Thanks for every answer.
Regards
Does the running front look like that at the end of the run? It might be possible that the current distribution within your gel is not even, i.e., because the chamber is not completely filled with running buffer.
It may be that one platinum wire electrode is broken. Check both anode and cathode wires to ensure that one is not broken and that the connection to the banana connectors is firm and secure
Did you concentrate the samples before loading? If you have different ion concentrations in your samples, the running front by be disturbed.
In the background of this image, I can see the power supply unit. Did you come across any error notification on that during the running process? If so, then you can figure out if there is a connection or any electrode related issue. Also, have you tried running it at a lower voltage like 80-100 for a longer duration?
It might be something related to the electrical current. The mobility of the ions, especially the protein-SDS ions relies on the Current. Thus, a constant current would result in a constant rate of migration. So, do check whatever is related to the electrical current (e.g., power supply, anode and cathode, etc.)
Hi
You can also try to prepare your gel cast yourself and try not to have air bubbles upon preparation
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