Protocols...
Category:
Year:
Keyword(s):
Field:
Keyword(s):
Field:
Keyword(s):
Field:
Welcome to LabWizards.com!

In order to access full contents on LabWizards.com, please log in or register now.
Email:


Password:


Forgot password?

Haven't you joined our community yet? Registration is free and will grant full access to website and will allow you to browse our protocols, upload original contents, create your own scientific portfolio and more.

Register now >>>

Created on: 25-Apr-2014. Updated on: 25-Apr-2014.

Category: Proteins.

Silver Staining Method Compatible with Mass Spectrometry

Posted by: Vania, Northwestern University, Chicago, IL



This Silver Staining Protocol was adapted from Shevchenko A et al, Anal Chem, 1996 and allows detection of proteins on a SDS-PAGE gel with high sensitivity but at the same time allows identification of the spots/bands by MS/MS analysis. 

Materials.
Fix solution: 5% acetic acid, 45% reagent alchol, 50% water
Sensitizer: , Silver and Developer reagents fresh every time. Fix 1: 50% methanol/10% acetic acid Fix 2: 5% methanol/1% acetic acid Fix 3: 50% methanol Stop: 6% acetic acid or 40 mM Na2-EDTA SENSITIZER: 0.02% sodium thiosulfate 20 mg Na2S2O3• 5H2O + 100 ml ddH2O Reserve a small volume of Sensitizer (2.0 mL) for Developer solution. SILVER: 0.2% silver nitrate 200 mg AgNO3 + 100 ml ddH2O Keep at 4ºC until used DEVELOPER: sodium carbonate/sodium thiosulfate/formaldehyde 6.0 g NaCO3 + 50 μl (37% stoc...

Login or register for accessing full contents