Good tips! To visualize lanes I also add a very tiny amount of bromophenol blue to the stacking gel mix to give the stack a light tinge of blue. A light blue stack makes seeing the wells easy. If I forget to do so I also sometimes rinse the wells with running buffer with very tiny amount of running buffer with bromophenol blue. It adds just enough contrast to see.
Amazing thanks for the tips!! I’m loading my gel today as my first real experiment. I was wondering, if I harvest 2 million cells instead of 1 million, lyse my cells and do my protein quantification, is it likely that I will have more protein conc in 2 million compared to 1 million? Trying to avoid loading a large volume in my wells!
If the volume is the same, it should be higher concentration. But beware that if you go too high in concentration you could overload the wells. Good luck!
i found myself here at 5 AM preparing to learn western blot tmrw :) thank you so much
Nice explanation, better than everything I have heard before.
Thanks! Glad it was helpful!
Super helpful, all the little unspoken tips in one place! Thank you for sharing.
Good job. This is very critical especially with loading sample from pellet!
Good tips! To visualize lanes I also add a very tiny amount of bromophenol blue to the stacking gel mix to give the stack a light tinge of blue. A light blue stack makes seeing the wells easy. If I forget to do so I also sometimes rinse the wells with running buffer with very tiny amount of running buffer with bromophenol blue. It adds just enough contrast to see.
Great tip!
Thank you for the valuable tips!
Glad it was helpful!
Amazing thanks for the tips!! I’m loading my gel today as my first real experiment. I was wondering, if I harvest 2 million cells instead of 1 million, lyse my cells and do my protein quantification, is it likely that I will have more protein conc in 2 million compared to 1 million? Trying to avoid loading a large volume in my wells!
If the volume is the same, it should be higher concentration. But beware that if you go too high in concentration you could overload the wells. Good luck!
Nice