Ponceau S Staining Solution
Chemical Name: 3-Hydroxy-4-[2-[2-sulfo-4-[2-(4-sulfophenyl)diazenyl]phenyl]diazenyl]-2,7-naphthalenedisulfonic acid tetrasodium salt
Biological ActivityStain used to detect protein bands after Western blotting.
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Protein samples were resolved in NuPAGE 4–12% Bis-Tris gel (Invitrogen) and transferred to 0.2 µm PVDF membrane. The membrane was stained with Ponceau S solution for less than a minute to detect protein bands and confirm successful sample run and transfer (lane 2-6) or not (lane 1).
We use Ponceau S (Cat. No. 5225) to confirm successful transfer of proteins from the gel onto nitrocellulose membranes during the transfer step of western blot analysis. This reagent stains protein bands very quickly (less than 1 min) and has the big advantage to be easily destained after a quick wash step that can be made with water or PBS. It is also used in our lab to detect protein degradation or any other abnormalities like overloading (as shown in the figure, the line #9 contains a protein sample degrade). Moreover, it is also useful to select the membrane area to be cut after transfer, based on the migration pattern of the protein standards (line #1). This enables one to incubate the different membrane sections with various primary antibodies.
Western blot confirmation