- November 9, 2009 at 7:53 pm #12241samkh918Participant
I stain my 2D gels with coomassie blue and sometimes about a week later, perform silver staining on them. I was wondering if the original coomassie blue staining has any effect on the quality of the subsequent silver staining.
I would also very appreciate if someone could send me a tested silver staining protocol.
Thanks a lot for your help.
- November 22, 2009 at 6:54 pm #95224ggsteamParticipant
I am not sure if the Coomasie staining interferes with the silver staining but it may be the case because both stains interact with the aminoacids of the protein. Additionally you need to fix the gel prior to the staining and I am not sure if the treatment of the gel with the Coomasie itself (maybe not if you use the colloidal Coomasie) may interfere with the fixing for the silver staining. I jus imagine it as putting two different stains on the same thing may not be a good idea. I am not sure if you will get the sensitivity you want if you use Coomasie and subsequently the silver staining. Additionally the Coomasie gives a high background which you probably do not want in your silver staining. If you need to stain with the Coomasie to be sure if you have any protein before you go with the silver I would first run the Coomasie with the samples you want to analyse and if it is ok then run another gel and stain it with the silver method.
Here you can find a protocols for the silver staining:
http://proteomics.cancer.dk/images/prot … aining.pdf
- November 28, 2009 at 9:36 pm #95462samkh918Participant
Thanks a lot!
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