Prior studies investigating misfolded SOD1 in ALS primarily relied on immunohistochemistry of spinal cord sections19,20,29,31,32, but this approach suffers from two major limitations: first, commonly used antibodies against misfolded SOD1 are inconsistent and often fail to detect all misfolded forms28; second, the high antibody concentrations required for staining make large-scale screening impractical19. This evidence concerns the gene SOD1 and amyotrophic lateral sclerosis.