Ponceau S is used for the staining of proteins immobilized on nitrocellulose filters (see references).
It is particularly suitable for reversible staining of proteins on transfer membranes during immunoblotting.
In a staining procedure using Acetic acid and Ponceau S proteins are fixated on the
membrane and proteins are stained reversibly. The Ponceau S stain is easy to remove.
For the Staining of Proteins During Immunoblotting
1. Unload the Nitrocellulose membrane from the blotting chamber
2. Wash briefly the membrane in buffer solution, e.g. TBST
3. Stain the transfer membrane in staining solution for 5-10 min at RT
4. Destain in water until the background becomes clean
5. Use a pencil to mark the positions of protein markers if wanted
The Staining of blotted proteins will disappear during subsequent
blocking. Alternatively, destain the blot in 5% acetic acid solution
for approximately 15 minutes
(1) Sambrook, J., Fritsch, E.F. & Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd Edition;
page 18.67. Cold Spring Harbor Laboratory Press, Cold SpringHarbor, New York.
(3) Bannur, S.V. et al. (1999) Anal. Biochem. 267, 382-389. Protein determination with Ponceau S on nitrocellulose
(4) Ausubel, F.A., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. & Struhl, K. (eds.) (1995) Current
Protocols in Molecular Biology, Page 10.8.7. (Suppl. 39), Greene Publishing & Wiley-Interscience, New York.
(5) Salinovich, O. & Montelaro, R.C. (1986) Anal. Biochem. 156, 341-347. Reversible Staining of Proteins Transferred
to Nitrocellulose after Separation by SDS-PAGE.