Understanding the iron accumulation process in ferritin protein nanocages has remained a centerpiece in the field of iron biochemistry/biomineralization, which ultimately has implications in health and diseases. Although mechanistic differences of iron acquisition and mineralization exist in the superfamily of ferritins, we describe the techniques that can be used to investigate the accumulation of iron in all the ferritin proteins by in vitro iron mineralization process. In this chapter, we report that the non-denaturing polyacrylamide gel electrophoresis coupled with Prussian blue staining (in-gel assay) can be useful to investigate the iron-loading efficiency in ferritin protein nanocage, by estimating the relative amount of iron incorporated inside it. Similarly, the absolute size of the iron mineral core and the amount of total iron accumulated inside its nanocavity can be determined by using transmission electron microscopy and spectrophotometry, respectively.
Keywords: Ferritin nanocage; Iron; Iron mineralization; Iron quantification; Iron staining; Native PAGE; TEM.
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.