Hey,
Has anyone ever used Hemin in cell cultures? specifically HEK 293 and hela cells (what conc. did you use?). I also wanted to know what I can dissolve the Hemin in and at what stock concentration. Any help would be great.

Thanks,
joe

We make 20 mM solution in PBS as stock. You need to read refs for working concentrations.

Thanks,
Also, is filter sterilization of the sock solution necessary?

Thanks for all your help

I meant stock =) not sock .. also shoudl the stock solution be stored at 4 C or -20C
Thanks
joe

So I tried dissolving Hemin in DMEM with 10% FBS at 10 mM. (did not filter since filtering seemed to remove all the hemin). It seems that I can't dissolve Hemin in the DMEM. I keep getting undissolved hemin when I spin down my solution, even after much vortexing. Any idea on how to prepare hemin to use in cell cultures. I added the above (undissolved) solution to my cells and I see large hemin particles under the microscope. Will these be absorbed by cells?
I'm waiting for DMSO, The lab does not have any at the moment.

Thanks.
joe

So i've just looked at my transfected cells. Cells incubated with 50uM Hemin for 48 hrs died. The Hemin was dissolved in DMSO at 1mM conc but not filtered. I tried filtering with a .2 micron filter but the DMSO solution seemed to dissolve the filter. Any other idea on how I can filter the stock solution. Anyone have an idea as to why the cells are dying? Is hemin too toxic at 50 uM or is it too toxic if incubated for 48 hrs.
whew working with a new compound is always so challenging

Thanks,
joe