The Thermo Scientific Pierce Silver Stain for Mass Spectrometry is a complete kit for rapid and ultra-sensitive silver staining of proteins in polyacrylamide gels and efficient destaining of excised gel pieces for mass spectrometry analysis.
This silver staining kit for mass spectrometry (MS) bundles components of the high-performance Pierce Silver Stain Kit with optimized reagents to destain spots for subsequent in-gel tryptic digestion to recover peptide fragments for proteomics analysis. The resulting MS-compatible product and protocol deliver outstanding sensitivity and maintains favorable conditions for the high-yield recovery and identification (sequence coverage) of proteins by mass spectrometry. Silver staining of 2D gels is now an important intermediate step in a set of procedures that leads ultimately to identification of specific proteins in the proteome by mass fingerprinting methods. The Pierce Silver Stain protocol provides peak staining performance, flexibility, reliability and robustness for applications such as MALDI-MS.
Sensitizer, stain, developer and enhancer solutions for silver staining gels; Reagent solutions A and B for destaining excised gel pieces.
Sensitive and fast staining – the low-background, easy-to-use silver stain provides subnanogram sensitivity, detecting proteins at less than 0.25ng per spot in 30 minutes after fixing
Flexible staining protocol – fix gel for 15 to 30 minutes or overnight without any affect on staining performance; stain for 1 to 30 minutes (typically 2 to 3 minutes)
Robust - effective silver stain for even difficult-to-stain basic proteins, such as lysozyme (pI 10) and chymotrypsinogen A (pI 9.2), which are detectable at 0.2ng and 0.5ng, respectively
Simple, trouble-free spot preparation – stained spots in excised gel-pieces are easily destained and made ready for tryptic digestion in one hour
MS compatible – reagents and protocol are optimized to provide excellent performance in mass spectrometry following extraction of peptides from stained 1D or 2D gels (SDS-PAGE)
Complete kit – contains all reagents necessary for staining and destaining before in-gel proteolysis and peptide recovery for analysis by mass spectrometry
Convenient – kit-components are stable at room temperature; saves valuable refrigerator space and eliminates the need to equilibrate reagents before use
The Pierce Silver Stain Kit for Mass Spectrometry enables both first-time and experienced users to achieve consistent and reliable staining using high-, low- and gradient-percentage polyacrylamide gels in single-dimension and 2D formats. The optimized staining method ensures extremely sensitive staining while minimizing covalent crosslinking of protein to the gel matrix, which can inhibit protein-peptide recovery following in-gel proteolysis. The destaining reagents facilitate complete removal of silver from stained protein bands and maximum protein recovery for subsequent mass spectrometry analysis.
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. However, the procedure accompanying the Kit for Mass Spectrometry is optimized to maximize polypeptide recovery without greatly sacrificing sensitivity.