Guidelines for the submission of samples
- First time users, non-affiliated researches and anyone wanting to submit a large number of samples (more than 15) are ask to contact the facility prior to sample submission.
- We highly recommend to use coomassie staining over silver staining. If you have to use silver staining, please make sure to use a mass spectrometry compatible staining protocol.
- Please provide an image of the gel and indicate, which band to analyze.
- Make sure that you clean your equipment thoroughly to avoid contaminations with keratins and other common contaminants.
Coomassie Stained Gels
We currently recommend to stain the gels with Imperial Stain™ (Pierce). This staining has the advantage that it uses no Methanol etc. which can lead to artifacts in the identification of the peptides.
- Destain the gel to get a clear background.
- Do not leave the gel for prolonged times in an acidic destain solution.
- If you cut out the gel bands yourself, please provide an electronic picture and indicate, which bands have been cut.
- Cut the bands with as little excess empty gel as possible.
- Place the bands in a clean centrifuge tube with a little drop of double destilled water.
Silver Stained Gels
- We strongly encourage to use Coomassie staining rather than silver staining.
- If you cannot visualize your bands with coomassie try to scale your preparation until you reach the limits of a coomassie brilliant blue staining.
- In case you cannot optain more material and you have to use silver staining, make sure that the protocol you use for the staining is mass spectrometry compatible. Do not use solutions, which contain formalehyde or glutaraldehyde for the fixing step. Refer to Shevchenko et al. (1996) Analytical Chemistry, 68:850-858 for more details or use this protocol.