About 10% of ALS cases are inherited; and a large subset of them are caused by mutations in the gene encoding the copper-zinc superoxide dismutase (SOD1). The detection of SOD1-positive inclusions in familial ALS patients suggests the role of SOD1 aggregation underlying the pathology of familial ALS. Although SOD1 mutant proteins are different in structure, stability and activity; they all exhibit a higher aggregation propensity than wild-type SOD1.
In this assay from Innoprot, we use a HEK293 cell line stably expressing the red fluorescent A4V mutant of SOD1; which is induced to form aggregates by incubating with ALLN, cysteine proteinase inhibitor. Before the aggregation process, cells are pre-incubated with the test compounds during 24 hours to evaluate their effect preventing SOD1-A4V aggregation. Then, the cells are treated with ALLN to induce aggreagation. After overnight incubation, the cells are fixed and the red fluorescent SOD1-A4V aggregates are quantified with an image analysis algorithm.
