I just add 1~2ng/ul FAC （Ferric Ammonium Citrate）into DMEM medium. perl's blue staining shows that cells have uptook the extra iron.

some papers also suggest transferrin+Fe3+ more efficient than FAC.

hm. iron biologically is transferrin-bounded and uptaked. most of the other heavy atoms do this (with other proteins, of course). but for your chelators, you need free toxic Fe - transferrin bounds strongly it. and moreover tc medium contains transferrin... so, check some relevant paper dealing with iron toxicity. citrate looks good for me.

Tnx Atilla Lehotzky

i found one paper that worked with FeSo4

how can i find normal range of interacellular Fe?

is feso4 toxic for cells?

heavy metals free concentration is low, somewhere in nanomolar range. Fe, Zn, Cu, Mn all are toxic, and cells deal them with special carriers, like transferrin, or other less specific ones, like methallothienins, small proteins. start to add your iron source to TC medium in wells with cells, increasing the concentration... first find a range of toxicity where cells still able to response by like MTT activation and live about 24 hours. then investigate your chelators in there range whether they lower toxicity of Fe. note that free Fe2+ quickly oxidized to 3+ in cellular environment.