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Thermo Scientific™ Pierce™ Silver Stain for Mass Spectrometry
Description
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.
Highlights:
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 effect 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
Includes:
Sensitizer, stain, developer and enhancer solutions for silver staining gels; Reagent solutions A and B for destaining excised gel pieces
Specifications
Specifications
| Content And Storage | Kit Contents: • Silver Stain Sensitizer, 2 mL • Silver Stain Enhancer, 25 mL • Silver Stain Developer, 500 mL • Silver Stain, 500 mL • Silver Destain Reagent A, 4 mL • Silver Destain Reagent B, 14 mL Storage: Upon receipt store at ambient temperature |
| Detection Location | In-Gel Detection |
| Detection Method | Colorimetric |
| Label or Dye | Silver |
| Quantity | 1 L |
| Target Molecule | Protein |
| Product Line | Pierce |
| Product Type | Silver Stain |
Frequently Asked Questions (FAQs)
Here are possible causes and solutions:
- Stained gel was overdeveloped. Reduce development time.
- Washing step(s) was missed or poor quality water was used. Do not skip or reduce wash steps and use ultrapure water.
- Contaminated equipment was used. Use clean equipment rinsed with ultrapure water.
- Impure chemical was used for gel preparation or precast gel has expired. Use analytical grade chemicals or use precast gels that have not expired.
- Stop solution was not effective in halting development of gel. Prepare new 5% acetic acid and replace it twice in the first minutes of incubation with the gel.
Here are possible causes and solutions:
- Insufficient development time. Develop gel for more than 5 minutes or add freshly prepared Developer Working Solution.
- Minimal or no protein present in sample. Check protein concentration in the original sample.
- Improper solution preparation or skipped steps. Check solution preparation and follow procedure.
- Excessive water wash before development step. Wash gel three times for 10 minutes each to completely remove the previous solutions and redo the staining procedure starting at step 2. Do not over wash prior to incubation in the developer.
After using the Thermo Scientific Silver Stain for Mass Spectrometry, proteins can be recovered and processed by several methods for mass spec analysis. For in-gel trypsin digestion, with or without reduction and alkylation, use the Thermo Scientific In-Gel Tryptic Digest Kit (Cat No. 89871). Consider following the recommendations of the core facility that will be performing the mass spec analysis.
The Thermo Scientific Silver Stain for Mass Spectrometry provides subnanogram sensitivity, detecting proteins at less than 0.25 ng per band in 30 mins after fixing.
We recommend storing the Thermo Scientific Silver Stain for Mass Spectrometry at room temperature where it is stable for a year.
For Research Use Only. Not for use in diagnostic procedures.