Hello everyone!
I forgot to take a picture of my Ponceau stain prior to starting my first round of primary and secondary antibodies. I'm currently doing my loading control now and was wondering if I could do the Ponceau stain again and take a picture for my records. Would there be any difference from the first staining I did?
Thank you for your help and feedback!
Dear Jasmine!
You look at following article, please:
Article Stain Free Total Protein Staining is a Superior Loading Cont...
After Stain free imaging of the gels, the gels were stained with ponceau S (Amresco, OH) for 1 minute and quickly destained in water to remove non-specific ponceau staining.
Dear Jasmine,
staining the membrane with Ponceau only makes sense before blocking has been done; Ponceau is a non-specific protein stain and will visualize all proteins including the ones used for blocking (e.g. BSA or casein). You will probably end up with a completely red-colored membrane and won't be able to discriminate between transferred proteins and proteins from the blocking solution.
No, you can't as Carsten Lange explained. On the bright side, reviewers don't like the Ponceau staining anyway if you did not include the actin or tubulin they will ask to include it and presenting Ponceau stain into the manuscript frequently invite criticism. I always do Ponceau stain to confirm my transfer and cut the unused parts of the membrane but never bother to take images. I always find it a bit strange as total protein staining supposed to be a more reliable housekeeper than housekeeping genes.
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