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Pierce Silver Stain Kit


Reliability and optimization via flexible fixing and staining steps.

Pierce Silver Stain Kit

The Thermo Scientific 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 with many 1D and 2D gel types.

The Pierce Silver Stain Kit is a metallic silver (Ag) protein stain that yields a remarkably clear and uniform gel background while delivering consistent, high-sensitivity staining results at each and every use. In standard mini gels, proteins are detectable at greater than 0.25ng 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.

Highlights:

  • Sensitive silver stain – detect proteins at 0.25ng; equal or better sensitivity than monochromatic metallic silver stains from other suppliers
  • Clear background – method produces intensely-stained protein gels with very low background color, shading or anomalies
  • Robust performance – reagents and protocol are formulated to yield excellent staining results amidst even wide-ranging differences in user technique
  • Rapid, flexible protocol – approx. 1.5 hours are required to complete the entire default procedure, but certain steps can be performed overnight or shortened with only minimal effects
  • Gel-compatible – 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

Additional Features:

  • Gel fixation step can be completed in 30 minutes or overnight
  • Gel staining can be performed within 5 min to 20 hrs, depending on desired sensitivity

Includes:

  • Sensitizer, stain, developer and enhancer solutions (1.03L total).

Product Details:

Thermo Scientific Pierce Silver Stain Protocol
Protocol summary for the Thermo Scientific Pierce Silver Stain Kit.

Careful manufacturing procedures and thorough quality testing ensure that the Pierce Silver Stain Kit yields excellent results with every use, independent of the polyacrylamide gel type or buffer system being used. Most proteins are easily detectable at low nanogram or subnanogram amounts, and background levels are very low (see Additional Information section at the end of these instructions). The protocol clearly indicates how both fixing and staining steps can be extended overnight without compromising performance, a feature that enhances convenience for ordinary workday schedules. Likewise, the default 30-minute staining step can be decreased to 5 minutes, enabling the staining procedure to be completed in less than 20 minutes after gel fixation.

Comparison of Pierce Silver Stain with Invitrogen SilverXpress Stain
Comparison of the Thermo Scientific Pierce Silver Stain Kit with Invitrogen SilverXpress* Stain. Dilutions of Escherichia coli lysate (1:25 to 1:1000) 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 Invitrogen™ SilverXpress™ Stain (Life Technologies). The molecular weight marker protein bands are myosin (14 to 0.2ng), phosphorylase B (55 to 0.9ng), BSA (14 to 0.2ng), ovalbumin (55 to 0.9ng) and carbonic anhydrase (55 to 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 has a flexible gel fixation step. The Thermo Scientific 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 to 0.2ng), phosphorylase B (55 to 0.9ng), BSA (14 to 0.2ng), ovalbumin (55 to 0.9ng), carbonic anhydrase (55 to 0.9ng), trypsin inhibitor (42.5 to 0.7ng), lysozyme (28 to 0.4ng), aprotinin (28 to 0.4ng) and insulin (42.5 to 0.7ng). 

 

The Pierce Silver Stain has a flexible staining protocol
The Thermo Scientific 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 to 0.2ng), phosphorylase B (55 to 0.9ng), BSA (14 to 0.2ng), ovalbumin (55 to 0.9ng), carbonic anhydrase (55 to 0.9ng), trypsin inhibitor (42.5 to 0.7ng), lysozyme (28 to 0.4ng), aprotinin (28 to 0.4ng) and insulin (42.5 to 0.7ng). 

 

Comparison of the Pierce Silver Stain and Invitrogen SilverXpress on a 2D Gel.
 
Comparison of identical 2D gels stained with Thermo Scientific Pierce Silver Stain Kit and another popular commercial silver stain formulation. Cytosolic protein extract (15µg) was separated by 2D 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 to 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.

 

Gel compatibility with the Thermo Scientific Pierce Silver Stain Kit.
Precast Gel Supplier Gel Buffer System Pierce Silver Stain
Performance Rating
Invitrogen™ Novex™ Gels
(Life Technologies)
Tris-Glycine
Bis-Tris
Tris-Tricine
Excellent
Excellent
Good
Thermo Scientific Precise Gels Tris-Glycine
Tris-HEPES
Excellent
Excellent
Bio-Rad™ Gels Tris-HCI
Tris-Tricine
Criterion (1D and 2D gels)
Good
Okay
Very Good
Homemade Gels 12% Tris-Glycine Excellent

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. The procedure accompanying the Pierce Silver Stain Kit for Mass Spectrometry (Part No. 24600) uses the same silver stain reagents but is slightly modified to optimize recovery without greatly sacrificing sensitivity.

Product 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.

Related Resources:

Pierce Silver Stain FAQ
Protein Gel Stains Selection Guide
Protein gel electrophoresis technical guide

Related Products:

Pierce Silver Stain Kit for Mass Spectrometry
Pierce Silver Stain Rescue Reagent


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
24612 Pierce Silver Stain Kit
Formulation: Proprietary solutions
Sufficient For: 20 mini gels (SDS-PAGE)

Kit Contents:
Silver Stain Sensitizer, 2mL
Silver Stain, 500mL
Silver Stain Enhancer, 25mL
Silver Stain Developer, 500mL
1L kit Product Instructions for product #24612 Pierce Silver Stain Kit MSDS for product #24612 Pierce Silver Stain Kit Certificate of Analysis for product #24612 Pierce Silver Stain Kit $131.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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