Enhanced Diagnostic Tools
Silver staining is less compatible with the mass spectrometric analysis. Therefore, we don't recommend using silver staining for any samples that will subsequently be submitted for MS analysis. If you can not visualize bands with coomasie and you are not able to scale up your isolation of protein to reach coomasie stainable levels, you must use a mass spec compatible silver stain protocol.
The following is a protocol used at the proteomics resource center for Silver staining. If you are using Silver staining kit from Sigma, Invitrogen, or other vendors, please follow their specific instructions.
For 1 gel, mix 75 ml methanol, 7.5 ml acetic acid, and 67.5 ml water.
0.02% Sodium Thiosulfate: For 1 gel, add 30mg sodium thiosulfate-5 hydrate to 150ml water.
0.1% Silver Nitrate and 0.08% Formalin (37%): For 1 gel, add 150 mg silver nitrate and 120ul 37% formalin to 150ml water.
2% sodium carbonate + 0.04% Formalin: For 1 gel, add 6g sodium carbonate to 300ml water. Just prior to use, add 120ul 37% Formaldehyde to the 300ml.
5% acetic acid: For 1 gel, add 7.5 ml acetic acid to 150ml water.
Preserving solution: For 1 gel, add 13.2ml glycerol (100% w/w) to 150 ml water.
If you have a query regarding any of our products or services,
please don't hesitate to submit your question below. A member of our support team
will review and answer your question as quickly as possible.