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!
blog: bit.ly/ponceau...
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: bit.ly/westernb... ; KZitem: • Western blots: the wha... & more practical lab tips and tricks here: bit.ly/lab_tri...
more about all sorts of things: #365DaysOfScience All (with topics listed) 👉 bit.ly/2OllAB0 or search blog: thebumblingbioc...
Негізгі бет Ponceau staining - what, why, & how with some practical tips
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