A Ponceau stain is a quick, easy, and reversible way to check your membrane after protein transfer in a western blot to get a sense of how things went. It helps you see if there are bubbles, whether you loaded even amounts, whether the lanes rain evenly, etc. It’s a total protein stain so it will show you all that’s there. It isn’t very sensitive but it does the trick!
YouTube: https://youtu.be/klnt9EGM6RI
basic protocol:
- run SDS-PAGE
- blot (transfer to membrane)
- rinse off in ddH2O (that really pure water) -> shake ~1 min
- pour on enough Ponceau solution to coat the membrane -> shake a couple min
- pour off Ponceau (can reuse lots of times)
- rinse background off with ddH2O
- take a picture
- wash all the way out with TBST
- block
- do your antibody probing stuff
more on western blotting: http://bit.ly/westernblotworkflow ; YouTube: https://youtu.be/Oun6_u6E090 & more practical lab tips and tricks here: https://bit.ly/lab_tricks_page
