How do you make a Ponceau S solution?

Preparation of ponceau S staining solution

1. For the preparing 500 ml of ponceau S staining solution add 25 ml of glacial acetic acid to 400 ml of distilled water.
2. Add 0.5 gm of ponceau S tetrasodium salt to the acetic acid prepared above.
3. Stir the mixture to dissolve.
4. Make up the solution to 500 ml using distilled water.

What is Ponceau S used for?

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.

When used on a membrane What does Ponceau S Do How is it able to do this?

Ponceau S is a negative stain, which binds to the positively charged amino groups of the protein and it also binds non-covalently to non-polar regions in the protein. Microgram quantities of transferred protein can be detected with Ponceau-S Stain, which generates reddish pink protein bands with a clear background.

Why would someone use Ponceau S instead of AB HRP color development or Coomassie?

Ponceau S staining protocol takes about 20 minutes, is non-toxic, and a gentler solution than Coomassie Brilliant Blue. Ponceau S staining solution does not fix the protein, allowing for western blot analysis after staining, which is another key consideration.

What is the protocol for Ponceau’s staining?

Protocol for ponceau S staining. Remove the membrane from the blotting chamber. Wash the membrane with TBS-T. Add the Ponceau staining solution till the membrane submerges. Keep it for gentle ration for 5-10 min at room temperature. Remove the staining solution (can be re-used) and add distilled water to remove the background.

How to make Ponceau’s stain with acetic acid?

Add 0.5 gm of ponceau S tetrasodium salt to the acetic acid prepared above. Stir the mixture to dissolve. Make up the solution to 500 ml using distilled water. Remove the membrane from the blotting chamber. Wash the membrane with TBS-T.

How to remove the background from Ponceau stain?

Remove the membrane from the blotting chamber. Wash the membrane with TBS-T. Add the Ponceau staining solution till the membrane submerges. Keep it for gentle ration for 5-10 min at room temperature. Remove the staining solution (can be re-used) and add distilled water to remove the background.

How long to use Ponceau’s stain on Western blot?

Following Western blotting, transfer the membrane to 5ml Ponceau S Stain solution. Place on an orbital shaker for 5 minutes at room temperature. Rinse membrane with DI water to achieve desired staining, approximately 2‐3 washes of 5 minutes each will remove the background staining.