|Pack size||1 kit (for 30 assays to stain the gel of 5 X8.5)|
|Application||To detect protein in polyacrylamide gels|
List of Components
|Silver Stain Sensitizer||1||30ml (100X)||AR0171-1|
|Silver Stain||1||30ml (100X)||AR0171-2|
|Silver Stain Developer A||1||1.5ml||AR0171-3|
|Silver Stain Developer B||1||120mlX5 (5X)||AR0171-4|
|Product Name||Silver Stain kit|
|Size||1 kit (for 30 assays to stain the gel of 5 X8.5)|
|Storage||Upon receipt store at 4°C. It is stable at 4°C for one year.|
|Label or Dye||Silver|
|Detection Location||In-Gel Detection|
|Equivalent||Thermofisher (Product No. 24612); Millipore Sigma (Product No. PROTSIL1)|
|Cite This Product||Silver Stain kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # AR0171)|
|Application||To detect protein in polyacrylamide gels
*Our Boster Guarantee covers the use of this product in the above tested applications.
Silver Staining Kit uses ammoniacal silver chemistry and glutaraldehyde sensitization to produce a highly sensitive silver stain, capable of detecting much lower levels of protein than standard Coomassie or Colloidal Blue techniques. Clear background makes sample identification unambiguous and provides publication-quality gels.
With the advantages of easy and effective operation, Boster's Silver Stain kit is suitable for the SDS-PAGE or non-denaturing PAGE. It is suitable for staining both single dimension SDS-PAGE gels and two-dimensional (2D) gels of complex protein solutions, and the staining gel will be compatible with subsequent mass spectrometry detection. The sensitivity of silver staining is 100 times as high as that of CBB (Coomassie brilliant blue) staining. The kit can detect 0.25 ng BSA. It is unnecessary to use the toxic methanol.
Boster's Silver Staining Kit is a highly sensitive silver staining kit providing nanogram-level sensitivity with minimal background. The easy-to-follow protocol requires less total time than standard Coomassie staining and is complete in 90 minutes.
Features of Protein Silver Stain Kit:
• Highly sensitive protein staining
• Crystal clear backgrounds
• A simplified protocol
• Ethanol and glacial acetic acid should be self-prepared.
• Avoid overstaining.
• Clean equipment should be used. Use gloved hands to manipulate the gel. When using gloved hands, touch the gel only at the edges to avoid depositing protein on the surface, which may cause background.
• Throughout the procedure, use sufficient volumes of solution to thoroughly cover the gel.
1. Immerse gel into 100ml Fixative, place it on a horizontal shaker with gentle shaking at 60-70rpm for 20 minutes.
Prepare Fixative by mixing 50ml ethanol with 10ml glacial acetic acid and 40ml deionized water.
2. Wash gel in 100ml of 30% ethanol. Shake at 60-70rpm for 10 minutes.
Prepare 30% ethanol by mixing 30ml ethanol with 70ml deionized water.
3. Wash gel in 100ml deionized water. Shake at 60-70rpm for 10 minutes.
4. Incubate gel in 100ml Silver Stain Sensitizer Working Solution (1X). Shake at 60-70rpm for 2 minutes.
Preapre Silver Stain Sensitizer Working Solution (1X) by mixing 1ml Silver Stain Sensitizer (100X) with 99ml deionized water. And use Silver Stain Sensitizer Working Solution (1X) within two hours after preparation.
5. Wash gel in 200ml deionized water and shake at 60-70rpm for 1 minute. Replace the water and shake at 60-70rpm for another 1 minute.
6. Incubate gel in 100ml Silver Stain Working Solution (1X). Shake at 60-70rpm for 20 minutes.
Preapre Silver Stain Working Solution (1X) by mixing 1ml Silver Stain (100X) with 99ml deionized water. And use Silver Stain Working Solution (1X) within two hours after preparation.
7. Wash gel in 100ml deionized water and shake at 60-70rpm for 30 seconds. Replace the water and shake at 60-70rpm for another 30 seconds.
8. Add 100ml Developer Working Solution and shake at 60-70rpm until protein bands appear (3-10 minutes).
Prepare Developer Working Solution by adding 20ml Silver stain Developer B (5X) into 80ml deionized water. Then add 0.05ml of Silver stain Developer A into the solution.
9. When the desired band intensity is reached, replace Developer Working Solution with prepared Stop Solution. Shake at 60-70rpm for 10 minutes.
Prepare stop solution by mixing 5ml glacial acetic acid with 95ml deionized water.
10. Replace Stop Solution with deionized water. Shake at 60-70rpm for 2-5 minutes.
Silver Stain kit Images
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