• Quickly determine the transfer efficiency of proteins on pure nitrocellulose/supported nitrocellulose and charged membranes
• Avoid re-doing western blots due to uneven transfer, non-transfer and air bubbles
• Stained membranes can be photographed for quality control step in the Western blotting protocol.
• Compatible with Nitrocellulose and PVDF membranes
• Preservatives allow for a shelf-life of 1 year at room temperature

StainODyeTM is a reversible and sensitive tool to detect transfer efficiencies of protein on nitrocellulose membranes. With fast response time, lower limit of detection of 25-50 ng per band and a quick and easy protocol StainODyeTM is a better alternative to traditional Ponceau S staining. Unlike other stains like Amido black/Coomassie, which permanently binds to proteins, StainODyeTM has a high affinity for protein, but is washed off in PBS or blocking solutions. Additionally StainODyeTM labels the glycosylated proteins with high avidity resulting visualization of total proteins.

Quantitative Western blotting applications requires uniform transfer of proteins to membrane for comparison analyses of results. There are accurate methods to load a fixed quantity of proteins on the gel before electrophoresis, however, few methods are available to quantitate the transfer efficiency of proteins on the nitrocellulose/charged membranes. Uneven transfer, non-transfer and air bubbles are some of the problem that can compromise quantitative western-blotting results. The StainODyeTM allows direct visualization of proteins on nitrocellulose or other types of transfer media immediately after transfer of proteins to charged membranes. The protein bands appear red or pink after membrane is developed in StainODyeTM. These stained membranes can be photographed for permanent records. The StainODyeTM is reversible and is washed off by rinsing the blots in water or PBS. Alternatively, they can be directly incubated in blocking buffer (BlockOBufferTM) for western blotting. The StainODyeTM comes with preservatives for an approximate shelf life of 1 year at room temperature.

1. Pour approximately 10-15 mL dye for 10 x 12 cm membranes in a clean glass container.
2. Incubate the membrane by slow shaking in a rocking platform for 1-2 minutes.
3. Remove and discard the excess dye and wash membranes with distilled water or PBS until the background is clear.
   1. Several washes may be necessary for optimal signal to noise ratios.
4. The membrane can be photographed immediately, dry membranes can be photographed by wetting in water.
5. There may be slight fainting of bands upon long-term storage. Best results are obtained by immediately recording the color intensity