A fast and more flexible silver stain with consistently low background.

The Thermo Scientific Pierce Silver Stain Kit is a more sensitive and reliable silver stain. The Pierce Silver Stain Kit (formerly called SilverSNAP Silver Stain Kit II) yields a remarkably clear and uniform gel background while delivering consistent staining performance each and every time. The protocol has been optimzed for flexibility by allowing short or overnight gel fixation and staining steps without causing negative affects on the results. In addition, 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.

Highlights of the Pierce Silver Stain Kit:

• Remarkably low, uniform background
• Detect as little as 0.25ng of protein
• Staining can be completed in a little as 50 minutes
• Option for shorter time, lower sensitivity protocol
• Optimal flexibility built into two key steps
  • Gel fixation step can be completed in 30 minutes or overnight
  • Gel staining can be performed within 5 min to 20 hrs depending on required sensitivity
• Compatible with elution for mass spectrometry

Thermo Scientific Pierce Silver Stain protocol.

Comparison of Pierce Silver Stain Kit with Invitrogen SilverExpress* Stain. Dilutions of Escherichia coli lysate (1:25-1:1,000) and dilutions of pure protein were separated by electrophoresis using 10% tris-glycine gels at 125 volts for 2 hours. The gels were then stained with Pierce Silver Stain Kit and the SilverXpress Stain. The molecular weight marker protein bands are myosin (14-0.2ng), phosphorylase B (55-0.9ng), BSA (14-0.2ng), ovalbumin (55-0.9ng) and carbonic anhydrase (55-0.9ng). The development times indicated below each gel image refer to the amount of time the gel was in the developer solution before reaching the desired level of staining and then stopped.

The Pierce Silver Stain protocol has a flexible gel fixation step. Dilutions of pure protein were separated by electrophoresis using 4-12% bis-tris gels at 200 volts for 35 minutes and stained with Pierce Silver Stain. The gel-fixing step (30% ethanol, 10% acetic acid) was performed for 30 minutes and overnight. The pure protein mixture preparation contains myosin (14-0.2ng), phosphorylase B (55-0.9ng), BSA (14-0.2ng), ovalbumin (55-0.9ng), carbonic anhydrase (55-0.9ng), trypsin inhibitor (42.5-0.7ng), lysozyme (28-0.4ng), aprotinin (28-0.4ng) and insulin (42.5-0.7ng).

The Pierce Silver Stain protocol has staining flexibility. Dilutions of pure protein were separated by electrophoresis using 4-12% bis-tris gels at 200 volts for 35 minutes. The gel-staining step was performed for 5 minutes, 30 minutes and overnight and then completed according to the protocol. The pure protein mixture preparation contains myosin (14-0.2ng), phosphorylase B (55-0.9ng), BSA (14-0.2ng), ovalbumin (55-0.9ng), carbonic anhydrase (55-0.9ng), trypsin inhibitor (42.5-0.7ng), lysozyme (28-0.4ng), aprotinin (28-0.4ng) and insulin (42.5-0.7ng).

Comparison of identical 2-D gels stained with Pierce Silver Stain Kit and a popular commercial silver stain formulation. Cytosolic protein extract (15µg) was separated by 2-D gel electrophoresis. The gels were stained with the Pierce Silver Stain Kit and another popular commercial silver stain according to the manufacturer’s recommendations. Both gels were developed for 1 minute and 40 seconds to reveal protein spots. The circled regions A- E on the respective gels indicate comparable regions of each gel and illustrate the differences in spot intensity and overall detection sensitivity. A side-by-side comparison clearly shows that the Pierce Silver Stain Kit was able to reveal spots that were faint or unobservable in the gel stained with the competing product.

Related Products and Resources

Pierce Silver Stain FAQ

References:

1. Koszelak Rosenblum, M.E., et al. (2002). J. Biol. Chem. 277, 11664-11669.
2. Mattagajasingh, S.N., et al. (2000). J. Biol. Chem. 275, 22728-22735.