Is there any method except autoradiography by which it can be identified that this unknown/known protein on the SDS-PAGE gel bind to a specific metal which could be Zn, Cu or As etc ?
A problem with using SDS-PAGE is that it denatures most proteins, which will cause any bound metals to dissociate.
If you have a sample of purified protein and you want to see if it has any metal bound, without using radioactivity, then you could submit it for atomic absorption spectroscopy or ICP-MS. Great care must be taken to avoid metal contamination that gives a high background, and any unbound metal must be removed.
Copper ions have a visible absorption spectrum, so if you have a sufficient quantity and concentration of the purified protein you should be able to see the color from the copper.
You could test a sample of purified protein for the presence of zinc(II) by denaturing the protein and adding a zinc(II)-dependent fluorophore.
Did you mean to write 'SDS-PAGE" or native gel? There are methods for native gels.
Thanks for reply . I meant to say native gel. It would be very nice of you if you could share few of those methods. I have gone through some of the research articles and come to know about the gel shift assay technique. If there are any others then please share?
Thank you so much for the reply Adam B Shapiro Sir and Grant Shimamoto Sir.
Hopefully you have the resources for this:
Talanta
Volume 76, Issue 5, 15 September 2008, Pages 1183–1188
Analysis of metal-binding proteins separated by non-denaturating gel electrophoresis using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS)
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