Hi,
I´ve been staining SDS page gels using the silver reduction technique. The main goal is to cut off differential bands (between treatments) from the gel and analyze it by MS. After purification process, I realized the WB bands (tagged) do not correlate with the silver pattern. Surprinsingly, some regions with WB signal match with "hollow" regions in the silver gel.
I´ve read about that mucins and negative-charged proteins do not stain with silver producing these "hollow" bands, but I must be able to visualize these proteins (and the others too..) in order to sequence.
Someone knows a protein detection technique to stain ALL the proteins with a sensitivity similar to silver staining?
Any other suggestions will be grateful
Best regards,
Martin
Have you tried fluorescent stains?
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2918041/
Try this will work.
http://camelot.bioc.cam.ac.uk/~marko/methods/zincimi.pdf
Periodic acid/Schiff will stain glycoproteins in gels and on blots, the latter is quicker and uses less chemicals.
@ARTICLE{Fai-71,
title = {Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane},
author = {G. Fairbanks and T.L. Steck and D.F.H. Wallach},
journal = {Biochemistry},
year = {1971},
pages = {2606-2617},
volume = {10},
doi = {10.1021/bi00789a030},
language = {eng},
}
@ARTICLE{Tho-89,
title = {Quantitation of mucus glycoproteins blotted onto nitrocellulose membranes},
journal = {Anal. Biochem.},
volume = {182},
number = {1},
pages = {160-164},
year = {1989},
doi = {10.1016/0003-2697(89)90735-5},
author = {D.J. Thornton and D.F. Holmes and J.K. Sheehan and I. Carlstedt},
}
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