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The membrane was incubated with rabbit anti-Glutathione Peroxidase 4/GPX4 antigen affinity purified polyclonal antibody (Catalog # A02059-1) at 0.5 g/mL overnight at 4C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT

A specific Mg 2+ /Zn 2+ ratio is necessary to avoid displacement of Mg 2+ and to obtain optimal activity

Over transferrin bound (TBI) and non-transferrin bound iron (NTBI) can overwhelm the heavy chain ferritin (FtH) capacity, which lead to release of labile iron and render proximal tubular epithelial cells (PTEC) susceptible to iron oxidant damage, especial in distal tubular epithelial cells (DTEC) with lack iron storage (light and heavy chain ferritin) and export protein (ferroportin) (48)

doi: 10.1111/1471-0528.16972 386 SobelJ