Run and Transfer your Gel
How to Run your gel will depend on your system. It’s important to run your gel enough to detect subtle differences.
There are some common problems when running your gel:
Smiling (curved) bands or blurred bands may indicate that the running was too fast, the buffer heated up or that you loaded too much quantity of protein. So, try to run at a lower voltage and to make sure your buffer keeps at Room Temperature during the whole running (we suggest to run it in a Cold Room) and/or load a little bit less of protein.
WB Transfer will also depend on your system. There are some very efficient systems that allow you to perform transfer of any size proteins in only 3 minutes!
In general, we can say that bigger is the protein, longer it takes to transfer it. One recommendation is to keep the transfer system at cold temperature. Longer transfers will need extra care with temperature issues. It is important to add an ice pocket and a stir bar to distribute the temperature along the system. If trying to transfer very big proteins, it is advised to transfer in a cold room (4ºC) and to replace the ice pocket after some time by a new one.
¿Looking for buffers preparation and recipe information?
Running Buffer 5x (1L):
Transfer Buffer 5x (1L):
(When preparing the 1x buffer, don’t forget to add 20% Methanol)