Amyloid-enhancing factor (AEF) is a biological “activity” that is defined in the context of inflammation-associated amyloidogenesis (AA). When administered intravenously to mice followed by an inflammatory stimulus, such primed mice deposit substantial AA amyloid in spleen within 36–48 h. Since experimental induction of AEF is dependent on amyloidogenic protocols, and rapid AA amyloid induction is dependent on AEF, a strategy for AEF isolation is required to break into this circular process. AEF activity may be prepared from a variety of human forms of amyloid that include tissue containing any of Aß, AA, ATTR, and AL amyloids. The preparation of an AEF extract from such human tissue is described using 4 M glycerol, which then may be used to induce splenic AA amyloid fibrils in mice as a source for the propagation of additional AEF and/or for the study of amyloidogenesis. The glycerol and AA fibril preparations are stable frozen for many years.