10X Concentrate Ponceaus S Protein Staining Solution is a 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 Protein 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 deionized water for 3 mins. It provides visible pink bands.

1. Prepare a 5L amber bottle. 
2. Unpack PS003-L10X and pour the liquid content into the bottle. 
3. Rinse residual reagent in the original bottle with deionized water. 
4. Add deionized water until the volume reaches 5L. 

1. Staining Solution. Make 10X concentrate Ponceau S Protein Staining Solution into 1X concentrate Ponceau S Protein.
2. After the SDS-PAGE Gel has been transferred to a nitrocellulose or PVDF membrane, place the transferred membrane in a staining container. 
3. Submerge the membrane in a proper amount of 1X 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. 
4. Wash away the excessive stain on the membrane with deionized water until the protein bands are clear. 
5.  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 deionized water for 1 to 2 minutes.