10X Concentrate Ponceau S Protein Staining Solution

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.

1. Prepare a 5 L 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 5 L.

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.

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.

• 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

Important notes

• Container: for membrane staining.
• Shaker: orbital or rocking shaker.