Membrane stains as close to ideal that you may never again use a Ponceau S membrane stain.

Thermo Scientific Pierce Reversible Protein Stain Kits for Membranes are high-performance alternatives to Ponceau S stain for detecting proteins on nitrocellulose and PVDF membranes. The Pierce Reversible Protein Stains have a detection limit of 25-50ng, produce stable turquoise bands which are easy to photograph and bands do not fade until the destaining process begins.

Highlights of Pierce Reversible Protein Stain Kits

• Sensitive, high-avidity, general protein stain
• Stain is protein-specific, avoiding interference from other biomolecules
• From stain to destain to band erasure in minutes
• Turquoise bands are easily photographed
• Stained bands do not fade with time
• All components are room temperature-stable

About Pierce Reversible Protein Stain Kits for Membranes

The Pierce Reversible Protein Stain Kits for Nitrocellulose and PVDF Membranes have improved avidity for a broad range of proteins, reducing protein-to-protein variability and increasing protein detection limits to 25-50ng of protein per band. In addition, the resulting turquoise-blue bands are easily photographed for documentation purposes and the bands do not fade until the destaining process begins. Staining is typically achieved with 30 seconds compared to 5 minutes with Ponceau S stain. Any background stain is easily removed from the membrane with a low pH wash and destaining of the bands completely removes the stain from the membrane allowing Western blot detection to be carried out with no adverse affects on target protein detection.

Traditionally, the red reversible stain Ponceau S was the best option for those staining Western blots to ensure complete protein transfer. Researchers who use Ponceau S for staining proteins are very familiar with its major shortcomings such as low sensitivity (~250ng), variable protein-to-protein staining and pale red bands which fade quickly and are difficult to document. The Pierce Reversible Protein Stain Kits resolve these issues.

Because of the different properties of nitrocellulose and PVDF membranes and their interaction with Pierce Reversible Protein Stain, two kits have been developed which are optimized for staining proteins on either nitrocellulose or PVDF membranes. It is important to note that these two kits, while they contain similar reagents, are not interchangeable for detecting proteins on nitrocellulose and PDVF membranes.

Thermo Scientific Pierce Reversible Protein Stain and Ponceau S stain. A comparison of GST lysate staining on nitrocellulose. Increasing amounts of GST Lysate protein were applied onto two 4-20% Tris-glycine SDS polyacrylamide gels. Proteins from both gels were electroblotted to nitrocellulose membrane. Panel A. was treated with Pierce Reversible Stain for 30 seconds and destained according to the protocol. Panel B. was stained with 0.1% Ponceau S stain for 5 minutes and destained. The blot stained with Pierce Reversible Stain demonstrates superior visual detection of bands. GST Lysate loading volumes (Lane 2-4). Lane 1. Thermo Scientific Pierce Prestained Marker Mix (Product # 26681), 10µL Lane 2. 15µL, Lane 3. 10µL and Lane 4. 5µL.

Comparison of Thermo Scientific Pierce Reversible Protein Stain with Ponceau S stain. Unstained Protein MW Markers were serially diluted and applied to two 4-20% Tris-glycine-SDS polyacrylamide gels (Lanes 2-10). Proteins from both gels were electroblotted to PVDF membrane. Panel A. Blot stained with Pierce Reversible Stain for 1 minute and destained according to the protocol. Panel B. Blot stained with 0.1% Ponceau S in 5% acetic acid for 5 minutes and destained according to the published protocol. Lane 1. Pierce Prestained MW Marker Mix (Product # 26681).

Immunoblot analysis of GST by chemiluminescent detection after treatement with Thermo Scientific Pierce Reversible Stain and destaining. Different amounts of purified GST protein were applied to two 10% Tris-glycine SDS-polyacrylamide gels. Proteins from both gels were electroblotted to nitrocellulose membranes. Panel A. Western blot was not treated with a reversible stain. Panel B. Western blot was subjected to the staining, destaining and stain erasing protocol of the Pierce Reversible Stain Kit. Both membranes were then probed with anti-GST incubated with goat anti-rabbit IgG-HRP conjugate (# 31460) and detected using Thermo Scientific SuperSignal West Dura Substrate (Product # 34075). Lane 1. 125pg, Lane 2. 250pg, Lane 3. 500pg and Lane 4. 1ng.

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