Immunoblot Analysis of Alzheimer's Proteins

Protein extracts were sonicated, centrifuged at 13,000 rpm for 45 min at 4^oC, and supernatants used for immunoblot analysis, as previously described (5 , 7 (link), 8 (link)). Actin was always used as an internal loading control. Primary antibodies used were as follows: anti GSAP full length (1:200) and GSAP-16kDa (1:150) (Thermo Scientific, Waltham, MA); anti-TMP21 (1:200) (Santa Cruz Biotech, Dallas, Texas); anti-APP (1:200), anti-APH-1 (1: 200) anti- CTF-α and anti-CTF-β(1:200) (Millipore, Billerica, MA, USA); anti-PS1 (1: 200), anti-Nicastrin (1: 200) (Cell signaling, Danvers, MA); anti-Pen-2 (1: 200) (Invitrogen, Carlsbad, CA); anti-β-actin (1:200) (Santa Cruz Biotech, Dallas, Texas). IRDye infrared secondary antibodies were from LI-COR Bioscience (Lincoln, Nebraska).