Ponceaus S Staining Solution is a ready-to-use membrane stain for evaluating the transfer efficiency of a western blot. This stain is recommended for rapid and reversible protein staining on nitrocellulose or PVDF membranes. Ponceau S staining is reversible and can be removed with a short incubation in 0.1% NaOH.

Figure 1 Ponceau S solution can be used to evaluate for total protein amount or transfer efficiency on nitrocellulose and PVDF membrane. The PVDF membrane stained Ponceau S for 5 mins and washed with ddH2O for 3 mins. It provides visible pink bands.

Lane 1, 13: Prestained Protein Ladder #PM019-0500

Lane 2-7: 2X Dilutions of Unstained Protein Ladder #PMU12-0500

Lane 8-12: 2000, 1000, 500, 100, 50 ng of BSA

• Container: box for gel staining
• Shaker: orbital or rocking shaker 

1. After the SDS-PAGE Gel has been transferred to a nitrocellulose or PVDF membrane, place the transferred membrane in a staining container.
2. Submerge the membrane in a proper amount of Ponceau S Protein Staining Solution that is enough to cover the whole membrane. Lightly agitate the staining container for 5 to 15 minutes at room temperature. Protein bands will be stained and start becoming visible in 5 to 15 minutes.
3. Wash away the excessive stain on the membrane with DI water until the protein bands are clear.
4. For the further membrane blocking step, remove the Ponceau S Staining Solution from the membrane by placing the membrane in 20 ml 0.1% NaOH for 1 to 2 minutes then rinse in DI water for 1 to 2 minutes.

• The Ponceau S Protein Staining Solution is provided as a ready-to-use solution, there is no need to dilute before use.