Altered Proteolysis in Fibroblasts of Alzheimer Patients with Predictive Implications for Subjects at Risk of Disease
Figure 1
The most representative typical and altered isoelectrophoretic (IEF) profiles of TK from cultured human fibroblasts. (a) Aliquots (30–40 μL; 1 mg/mL) of fibroblast extracts were prepared and separated by IEF within a pH range of 3–10 and then blots were probed with anti-human TK antibody (see Section 2 and [25]). Negative samples presented no distinct band or just a faint signal over pH 8.4, while positive samples exhibited one to three heavily stained TK-ab isoforms migrating toward the alkaline region of gel (pH range of 8.5–9.4). (b) Densitometric analysis of immunostained TK was carried out with the aid of the ImageJ software and values of intensity were expressed as arbitrary units; the ratio between values of TK-ab (pH ≥ 8.5) and those of the normal TK band (pH = 8.4) for each sample is reported. The border between negative and positive samples was arbitrarily set at a ratio of 0.4. TK, transketolase.
