Anti human cd63

Primary antibodies utilized include anti‐human PLAP (Abcam, SP‐15, ab16695, rabbit monoclonal [WB 1:1000]); biotinylated anti‐human PLAP (Thermo Fisher Scientific, 8B6.18, MA512691); anti‐humanCD63 (Santa Cruz Biotechnology, E‐12, sc‐365604, mouse monoclonal [WB 1:1000]); anti‐human ALIX (Thermo Fisher Scientific, PA5‐52873, rabbit polyclonal [WB 1:1000]); anti‐human TSG101 (Thermo Fisher Scientific, JJ0900, MA5‐32463, rabbit monoclonal [WB 1:5000]); anti‐human apolipoprotein B (R&D, AF3556, polyclonal goat antibody [WB 1:2000), and rabbit polyclonal anti‐human Calnexin (Novus Biologicals, Littleton, Colorado, USA, NB100‐1965 [WB 1:500]. Secondary antibodies were obtained from the Jackson Laboratory and included horseradish peroxidase (HRP)‐conjugated to goat anti‐rabbit IgG (111‐035‐144, [WB 1:3000]), and HRP‐conjugated to goat anti‐mouse IgG (115‐035‐146, [WB 1:3000]).

Five percent agarose (Sigma-Aldrich) was dissolved in a buffer containing 25 mmol/L Tris base, 190 mmol/L glycine and 0.1% sodium-duodecyl-sulfate (SDS), and a horizontal gel was prepared. Optiprep^TM gradient fractions with identical volumes were pelleted, resuspended in lysis buffer, and electrophoresed in agarose-SDS gel at constant 100 V for 5 h. Proteins were blotted to PVDF membranes (Bio-Rad). Membranes were blocked in 1% BSA in TBS-Tween, and were incubated overnight at 4 °C in the same buffer with rabbit polyclonal anti-human-apoB100/48 (Novus Biologicals), or with rabbit monoclonal anti-human-CD63 (Santa Cruz Biotech). Secondary immunostaining was performed with a goat anti-rabbit-immunoglobulin-HRP conjugate (Abcam). Chemiluminescent signals were detected with the Pierce ECL Western blotting substrate (Thermo Fisher Scientific). Blots are shown in Supplementary Figs S12 and S13.