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Dementia diagnosis is associated with changes in antidiabetic drug prescription: an open-cohort study of ~130,000 Swedish subjects over 14 years

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In brief, samples were kept at 20 C for 60 min to allow protein precipitation

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The PVDF membrane was subsequently washed (5 min 5) with tris-buffered saline with tween (TBST) buffer and then incubated with the appropriate secondary antibody (Peroxidase-conjugated goat anti-rabbit IgG (H + L), at a dilution of 1:5000) in a 5% bovine serum albumin solution for 1 h at 25 C

This is a big shoe to drop from a validation standpoint