Tips for loading PAGE gels - the strategy that works for me

[@ritheshvooturi2310]

i found myself here at 5 AM preparing to learn western blot tmrw :) thank you so much

[@Jenny-ym5rg]

Super helpful, all the little unspoken tips in one place! Thank you for sharing.

[@steffighter144]

Nice explanation, better than everything I have heard before.

[@thebumblingbiochemist]

Thanks! Glad it was helpful!

[@MohammedAli-bj9jk]

Good job. This is very critical especially with loading sample from pellet!

[@fruitflylikeabanana]

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.

[@thebumblingbiochemist]

Great tip!

[@MA-yt4xf]

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!

[@thebumblingbiochemist]

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!

[@eduardoduarte1968]

Thank you for the valuable tips!

[@thebumblingbiochemist]

Glad it was helpful!

[@whodevrajverma]

Nice