Titration of BSA visualized by Ponceau S Staining Solution after polyacrylamide gel electrophoresis and transfer to nitrocellulose membrane.
1. Following protein electrotransfer, rinse membrane briefly in ddH2O to remove any detergent that may inhibit staining.
2. Incubate membrane in Ponceau S Staining Solution for 5-10 minutes at room temperature.
3. Wash membrane in ddH2O until distinct reddish-pink protein bands are visible (1-5 min).
4. If desired, mark proteins with waterproof ink or a pencil. Alternatively, a photograph of the stained membrane can also be taken.
5. Wash membrane in 1X TBS-T several times for 5 min at room temperature until protein bands are no longer visible.
6. Proceed with blocking step of Western blot procedure.
Store at room temperature. This product is stable for 12 months. Do not dilute or aliquot.
Ponceau S Staining Solution is supplied as ready to use. This product is recommended for rapid and reversible protein staining on nitrocellulose or PVDF membranes. This staining technique is often utilized to confirm protein electrotransfer in Western blotting assays prior to antibody-based detection.
Ponceau S is a negatively charged, red colored stain which binds to positively charged amino groups and non-polar regions of proteins. It has a detection limit of around 250 nanograms of protein following SDS-PAGE and electrotransfer to nitrocellulose membranes (1).
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