Advantages at a glance
- Stains proteins reversible
- Composition: 0.1% Ponceau S in 5% acetic acid (w/v)
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Product information "Ponceau S Solution"
Ponceau S is used for reversible staining proteins blotted on nitrocellulose or PVDF membrans and while running a TAU electrophoresis. The reversibility of the staining process is ideally suited for subsequent immunoblots (western blot) or other colour reactions as Ponceau S can be completely removed. Ponceau S is not stable for long time, thus proteins and protein markers should be marked with a pencil. For staining the membrane (nitrocellulose or PVDF) is incubated in a 0.1% - 0.5% Ponceau S solution in 5% acetic acid (alternatively in 3% trichloracetic acid) for 5 - 10 minutes. Destaining is done by incubating the stained membrane in 5% acetic acid for 15 minutes. Our Ponceau S solution is offered as 0.1% Ponceau S (w/v) in 5% acetic acid (v/v).
Specifications:
Composition: 0.1% Ponceau S in 5% acetic acid (w/v)
Density (d 20°/4°): 1.005
n 20°/D: 1.338
Application:
for reversible staining of blotted membranesSource:
syntheticSafety Information:
Classification:
Documents:
Safety Data SheetCertificate
Category List
After proteins have been separated by gel electrophoresis (SDS-PAGE) and transferred onto a membrane, the result of the transfer is initially not visible, making it difficult to assess the success of the blotting procedure and to perform subsequent steps reliably.
Reversible staining of proteins with Ponceau S can help overcome this problem, for example in the following applications:
1. Verification of transfer efficiency (blotting from a polyacrylamide gel onto a blotting membrane)
Ponceau S staining immediately reveals whether the proteins have been transferred evenly and completely from the gel onto the membrane. It also shows whether air bubbles or uneven pressure have interfered with the blotting process. Such transfer defects appear as unstained spots or “holes” within the protein bands.
2. Loading control and normalization
Verzeroli C. et al. demonstrated that there is a very good correlation between the amount of protein and the staining intensity obtained with Ponceau S (Verzeroli C. et al. (2023): A fluorescent Ponceau S-based total protein normalization method for conventional and challenging immunoblot samples. Anal Biochem, 681:115330.).
Based on this finding, the amount of protein in the individual separated bands can, at least approximately, be estimated.
3. Total protein normalization
In modern quantitative Western blot analysis, Ponceau S staining is increasingly used to compare the band (signal) intensity of target proteins with the total amount of protein loaded, or to calculate the proportion of target protein relative to the initial total protein amount. (See also Verzeroli C. et al.)
4. Orientation aid / marking of protein bands
Since Ponceau S also makes unstained molecular weight markers (protein ladders) visible, the positions of the molecular weight standards can be precisely marked on the membrane with a pencil. These markings can subsequently serve as a reference for determining protein size when a specific detection of the target protein is performed on the unstained membrane using antibodies.
5. Cutting the blotting membrane
Visible protein bands make it easier to accurately cut the membrane into strips so that the individual strips can be incubated in parallel with different antibodies.