Ponceau S stain is often used to evaluate the transfer efficiency of western blotting. It reversibly stains the protein on the membrane (nitrocellulose and PVDF membrane). So after checking the transfer by Ponceau S staining, the membrane is destained by washing with TBST and is used for immunodetection.
A 0.1% (w/v) solution of Ponceau S in 5% acetic acid is used for membrane staining. Here we describe a procedure to prepare 100 ml of Ponceau S staining solution.
Reagents and solutions
Glacial acetic acid
Equipment and disposables
250 ml bottle with a lid (e.g., Duran® laboratory bottles, with caps Sigma-Aldrich)
Magnetic stirrer and magnetic bead (optional)
0.1% (w/v) Ponceau S
5% acetic acid
Preparation of 100 ml of Ponceau S Staining Solution
Step 1: To prepare 100 ml of staining solution, weigh out 0.1 g Ponceau S stain in a 250 ml bottle.
Wear a lab coat and gloves while handling Ponceau S stain.
Step 2: Prepare a 5% acetic acid solution in water by adding 5 ml glacial acetic acid to 95 ml of deionized/distilled water.
Step 3: Add 100 ml 5% acetic acid (step 2) solution to a 250 ml bottle and shake/swirl the bottle until all Ponceau S dissolves completely.
The solution can be stored at room temperature.