Generation of polyclonal antibodies
The ab338 rabbit antibody was raised against a human Aβ21–34-peptide which had been conjugated to keyhole limpet hemocyanin (KLH) with maleimide reagent via its N-terminal cysteine (KLH-Cys-Aβ21–34; NH2-KLH-C-AEDVGSNKGAIIGL-COOH). The rabbit was immunized with a 1:1 (v/v) mixture of immunogen (100–200 µg) and Freund’s complete adjuvant and vaccinated three times with Freund’s Incomplete Adjuvant. The serum was purified by affinity chomatography against the Aβ21–34-peptide immobilized on an antigen-coupled sepharose column that subsequently was washed and eluted with 0.1 M glycine–HCl (pH 2.5) followed by rapid neutralization. This was all done at Agrisera (Agrisera, Vännäs, Sweden, ethical permit id A146–12).
Indirect Aβ ELISA
MaxiSorp plates (Nunc, ThermoFischer, Waltham, MA, USA) were coated overnight (o.n.) with…
