Staining PVDF Membrane with Ponceau S after Protein Transfer


Like nitrocellulose membrane, PVDF membrane can also be stained with Ponceau S stain to reveal protein bands after western blotting. However, staining is not as efficient and takes bit longer time to stain PVDF membrane. Stained PVDF membrane can be destained by washing with buffer (TBST/PBST)  and can be used for immunodetection of specific proteins.


Reagents and solutions
0.1% (w/v) solution of Ponceau S
PVDF membrane with proteins
Deionized/distilled water

Equipment and disposables
Container that can accommodate nitrocellulose membrane


Step 1: Remove the PVDF membrane after transferring the protein from the polyacrylamide gel. Now keep the membrane in Ponceau staining solution submerged at room temperature.

Step 2: Shake gently for 15 min. Remove the stain and rinse the membrane with Distilled or Deionized water two to three times.

Note: Protein bands will be seen as red/pink color bands. Take a photograph for the record.

Step 3: Destain the membrane by washing with PBST/TBST several times until all pink color disappears. Usually, it takes 2 – 3 washings.

Now process the membrane for immunodetection.

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