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Ponceau S Solution

Ponceau S Solution allows for the rapid and reversible detection of proteins on nitrocellulose and PVDF membranes for the verification of the transfer efficiency of Western Blotting before proceeding with incubation with primary antibody.

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Description

Ponceau S Solution allows for the rapid and reversible detection of proteins on nitrocellulose and PVDF membranes for the verification of the transfer efficiency of Western Blotting before proceeding with incubation with primary antibody. Transfer artifacts, such as uneven transfer or bubbles, can be quickly detected, preventing continuation with imperfect membranes, saving you time and expensive antibodies. Ponceau S does not damage the antigen or has any deleterious effects on the sequence of blotted proteins and polypeptides and is therefore the method of choice for location for blot?sequencing. Staining can be carried out in less than 5 minutes and destaining also takes only a few minutes of incubation in water. After destaining, one can proceed immediately with subsequent steps of the Western Blotting protocol.

 

Additional information

Quantidade

500 mL

Features

Product: ready-to-use Ponceau S solution for the reversible staining of proteins blotted onto nitrocellulose or PVDF membranes.
Quantity: 500ml
Storage: 2ºC – 25ºC for at least 1 year.

Documentos

Product Info – Ponceau S Solution

Protocols

1. After Western Blotting, image the blot for permanent record, and then rinse the blot with 20ml of water for 5 minutes at room temperature, using an orbital shaker.
2. Decant and incubate the membrane in a container containing 10-20ml of GRS Stripping Buffer (membrane must be completely covered) at room temperature for 5-10 minutes, using an orbital shaker
3. After stripping, wash the membrane extensively (min 3x) with wash buffer (e.g. TBS(T) or PBS(T)).
4. After washing, block the membrane as usual.
5. After blocking the membrane is ready for the next detection with other set of antibodies.

Note: Because Stripping may reduce signal intensity, it is recommended to probe first for the antigen with the lowest level of expression. If more stringent stripping is required, one could add reducing agent (DTT or β-mercaptoethanol) to the stripping buffer and/or heat the blot during the stripping procedure.

Order Information

Reference Description Quantity
GB12.0110 TBE Buffer 1L 10X
GB12.0510 TBE Buffer 1L 10X
     
     
     
     
     

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Product: Ponceau S Solution

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FAQs

How to use Ponceau S Solution:
Short Protocol
1. After Protein Transfer, wash the blot with plenteous pure water, for 10 minutes.
2. Immerse the blot in 5?20ml of Ponceau S Solution (>0,2ml/cm2 membrane), and stain for 5 minutes, using an orbital shaker
3. Decant the protein staining solution and rinse the membrane with sufficient amounts of pure water, until the background is reduced. Prolonged rinsing might lead to destaining of protein bands.
4. Image the blot for permanent record and destain the blot in pure water for 10?20 minutes, or alternatively with 0.1M NaOH, using an orbital shaker. Note that complete destaining is only achieved in the subsequent blocking step and that the Ponceau S stain will not interfere with either blocking or antibody binding steps.
5. Continue with the Western Blotting procedure as normal.


Bibliography
Gallagher, S. et al.(2008) Immunoblotting and immunodetection. In: Curr Protoc Mol Biol 83:III 10.8.1-10.9.28