I am working on NFU3 protein, an scaffold Chloroplast target protein with CXXC motif. We believe that this protein involved in Fe-S clustering and their assembly. I am looking for the nice protocol to perform a Fe-S assembly assay. It will be highly appreciable if some one suggested a good protocol.
I worked with human Fe-S protein. You must work in anaerobic conditions (glovebox).
Hi Krishna,
Yes, as Diego also mentioned, Fe-S cluster reconstitution is carried out under strictly anaerobic conditions inside a Vacuum Atmospheres glovebox under an inert atmosphere (
Dear Diego and Rodgoun,
Thank you so much for suggestions. In fact at the moment I am just looking which Protocol will be fit to my experiment, since we do not know whether NFU3 bid to teh Fe-S directly or we need to purify it? I am looking some direct method which can show whether NFU3 involves in Fe-S clustering or not. I hope I will have couple of choice to perform this killer experiment.
Have a great day,
Krishna
Hi Krishna,
One of the leading experts in this field is Prof. Caryn Outten. She has published numerous articles on proteins that bind Fe-S clusters, mostly in collaboration with Dr. Johnson ( as you will notice in their publications). I am sure you will find a killer protocol in her publications.
Good luck and let me know if you needed help with finding references.
R
Thank you so much Rodgoun,
I will definitely let you know, if I need some thing more related with this experiment.
Thanking you,
Kris
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