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Hereafter, the medium was removed, tissue 4% PFA-fixed for 20 min, washed twice with excess PBS and permeabilized by incubating in PBS–0.1% Triton X-100–2% BSA for 1 h

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In humans the ALAD molecule appears to foster competition between Zn2+ and Pb2+ for the same binding site,[29–31] since these zinc ions can be replaced by lead, leading to inactivation of the ALAD enzyme during lead poisoning.[32] ALAD-deficiency porphyria is an inherited disease and is associated with various natural variants of human ALAD.[29–31] The mutations that perturb the ALAD gene collectively destabilize the tertiary and quaternary structure of the enzyme, resulting in a reduction of the enzyme’s half-life as well as preventing proper folding