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3 protocols using anti caveolin 2

1

Immunoblot analysis of Caveolin proteins

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Immunoblots were analyzed as described elsewhere [21 (link)]. Detergent-soluble protein fractions were prepared from Caco-2 monolayers incubated with TW or controls and detected by the primary monoclonal anti-Caveolin-1 (Sigma-Aldrich), anti-Caveolin 2 (BD Pharmingen, Heidelberg), anti-rab5, anti-rab7, and anti-cathepsin (Cell Signaling, Cambridge), and anti-TW antibodies.
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2

Analyzing Alzheimer's Disease Pathways

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All chemicals and reagents were purchased from Sigma–Aldrich (Poole, UK), Fisher Scientific (Leicester, UK) or Vector Laboratories Ltd (Peterborough, UK) unless specified. Antibodies used were: anti N-APP (22C11; WB 1:500, IHC 1:20) and total and phospho anti-GSK-3α/β (1:750) (Millipore, Watford, UK); anti-Aβ (6E10, BioLegend, San Diego, CA, USA); anti-clathrin heavy chain (CHC) (Clone 23; WB 1:1200, IHC 1;20), anti-caveolin-2 (Clone 65; WB 1:200, IHC 1:50), anti-flotillin-1 (Clone 18; 1:300) and anti-dynamin-2 (1:500) (BD Biosciences, Oxford, UK); anti-BACE1 (1:500, Merck Chemicals Ltd. Nottingham, UK); anti-AP180 (LP2D11; 1:750), anti-PICALM (WB 1:700, IHC 1:100), anti-flotillin-2 (1:500) and anti-AP-2 (1:500) (Novus Biologicals, Littleton, CO, USA); anti-caveolin-1 (WB 1:800, IHC 1:100, Cell Signalling Technology, Beverly, MA, USA); anti-caveolin-3 (1:200), anti-presenilin-1 (PS-1, 1:250), anti-ADAM10 (1:5000), anti-SORLA (EPR14670; 1:400), anti-dynamin-1 (1:300) and anti-BIN-1 (1:100) (Abcam, Cambridge, MA); anti-Rab5 (S-19; WB 1:300, IHC 1:30, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and total tau (1:2000, Dako, Hamburg, Germany). Monoclonal anti-phospho-tau Ser396/Ser404 (PHF-1, 1:100) was a generous gift from Prof. Peter Davies, Albert Einstein College of Medicine, Bronx, NY, USA.
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3

Proteomic Analysis of Lipid Rafts

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The following primary antibodies (Abs) were used: anti-caveoin-1 (BD bioscience, Franklin Lakes, NJ; Abcam, Cambridge, MA), anti-caveolin-2 (BD bioscience), anti-flotillin-1 (BD bioscience, Franklin Lakes), anti-flotillin-2 (BD bioscience, Franklin Lakes), anti-CD71 (transferrin receptor, Santa Cruz Biotechnology, Santa Cruz, CA), anti-parkin (Santa Cruz Biotechnology, Santa Cruz, CA), anti-flag (Sigma-Aldrich, St. Louis, MO) and biotin conjugated anti-CTxB (Sigma-Aldrich, St. Louis, MO), anti-myc (Sigma-Aldrich, St. Louis, MO) and anti-actin (Santa Cruz Biotechnology, Santa Cruz, CA). MG-132 and lactacystin were purchased from Sigma-Aldrich (St. Louis, MO). Cycloheximide was purchased from Calbiochem (Denvers, MA). Rhodamine-conjugated transferrin and BOIPY® FL C5-Lactosylceramide were purchased from Invitrogen/Molecular Probes (San Dieogo, CA).
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