Pierce™ Silver Stain Kit

The Pierce Silver Stain Kit is a rapid, ultra-sensitive, and versatile silver stain system for protein detection in polyacrylamide gels, yielding consistent and reliable results. It is a metallic silver (Ag) protein stain that yields a remarkably clear and uniform gel background. In standard mini gels, proteins are detectable at greater than 0.25 ng per band or spot. The protocol has been optimized for flexibility by allowing short or overnight gel fixation and staining steps without affecting staining performance (sensitivity or clarity). A short (one-minute) sensitization step, performed after gel fixation, yields results that are free of the characteristically dark or blotchy backgrounds often seen with homemade or other commercially available silver stains. This feature is beneficial when performing densitometric analysis of silver stained gels.

Features:
• Sensitivity—detect proteins at 0.25 ng
• Protocol—gel staining can be performed within 5 min to 20 hrs, depending on desired sensitivity
• Gel-compatibility—staining performance is very good with a variety of commercial precast mini-gels and SDS-PAGE buffer-systems
• Workflow-compatible—mild chemical formulation ensures compatibility with mass spectrometry, sequencing, and other downstream procedures that depend on destaining and protein recovery

Silver staining methods generally use either glutaraldehyde or formaldehyde, which cause some covalent crosslinking of protein to each other and the gel matrix. To the extent that this crosslinking occurs, extraction or elution of protein from the gel will be inhibited. Pierce Silver Stain uses formaldehyde in the stain and developer working solutions, but the concentration is low enough that irreversible protein crosslinking is minimized.

FAQ

Can nucleic acids be detected with the Thermo Scientific Silver Stain Kit?
Yes. References in the literature and molecular biology protocols indicate that nucleic acids can be detected in polyacrylamide gels with silver stains of this type.

Can a Coomassie-stained gel be secondarily stained with the Thermo Scientific Silver Stain Kit?
Yes. First destain the Coomassie-stained gel to completely remove background. If an acid or methanol destaining solution was used, thoroughly wash the gel with ultrapure water, then proceed with the Thermo Scientific Silver Stain Kit protocol.

Can gels be dried after staining with the Thermo Scientific Silver Stain Kit?
Yes. Thoroughly wash the gel with water to remove the acetic acid and then soak the gel in a solution containing 5% glycerol and 10% methanol (or 10% ethanol) for one hour before drying in a gel-drying apparatus.

Can I process gels stained with Thermo Scientific Silver Stain Kit for mass spectrometry analysis?
Yes, see for the procedure for processing gels stained with Thermo Scientific Silver Stain, for mass spectrometry analysis. However, for best results, we recommend using the Thermo Scientific Silver Stain for Mass Spectrometry (Cat. No. 24600). This kit includes an optimized procedure and all the necessary reagents for staining gels and destaining gel pieces before in-gel trypsin digestion and mass spectrometry.

Can the bands be removed and analyzed by mass spectrometry following staining when using the Thermo Scientific Silver Stain Kit?
Yes. See Tech Tip: Process Stained Polyacrylamide Gels for Mass Spectrometry. Alternatively, use our Thermo Scientific Silver Stain for MS (Cat No. 24600), which is the same silver stain kit but also includes destaining reagents and a protocol optimized for maximum protein recovery.