Mouse monoclonal anti tsg101

Equivalent amounts of cells and EV protein lysates were electrophoresed and transferred to nitrocellulose membranes. Nitrocellulose membranes were then blocked in 5% non-fat milk or 5% BSA, 10 mmol/l Tris-HCl pH 7.5, 100 mmol/l NaCl, 0.1% Tween-20, and probed with the following primary antibodies: mouse monoclonal anti-GM130 (1:1000, BD Biosciences, 610822), mouse monoclonal anti-Annexin V (1:500, Santa Cruz Biotechnology, Dallas, U.S., sc-74438), mouse monoclonal anti-Annexin XI (1:500, Genetex, CA, U.S., GTX33010), mouse monoclonal anti-TSG101 (1:500, Santa Cruz Biotechnology, sc-7964) rabbit monoclonal anti-TERT (1:500 1000 Rockland Immunochemicals Inc., PA, U.S., 600-401-252S), mouse monoclonal anti-Pro-COL1A2 (D-6) (1:200, Santa Cruz Biotechnology, sc-166572), rabbit monoclonal anti-Integrin beta1 (1:2000, ab179471), rabbit monoclonal anti-MMP3 (1:1000, ab52915), and rabbit monoclonal anti-MMP14 (1:2000, ab51074) (all purchased by Abcam) and incubated in the presence of specific, HRP-conjugated secondary antibodies. Immunoreactive bands were visualized using ECL detection system (Amersham International, L.C., UK) and by ChemiDoc XRS Imaging system (BioRad, CA, U.S.) or G:Box Chemi XT Imaging system (Syngene, Cambridge, UK).

The following antibodies were used in this study: mouse monoclonal anti-TBEV E (United States Army Medical Research, Institute of Infectious Diseases, Fort Detrick, Frederick, MD, USA), mouse monoclonal anti-TBEV prM (United States Army Medical Research), mouse monoclonal anti-TSG101 (Santa Cruz), mouse J2 anti-dsRNA (SCICONS, Hungary), mouse monoclonal anti-ALIX (Santa Cruz), mouse monoclonal anti-CHMP4A (Santa Cruz), rabbit anti-TSG101 (Atlas antibodies), rabbit anti-CHMP4A (Invitrogen), rabbit anti-ALIX (Atlas antibodies), rabbit anti-HA tag (Invitrogen), mouse anti-HA tag (Invitrogen), rapid anti-GRP94 (Invitrogen), mouse anti-GAPDH (Sigma), mouse anti-βactin (Sigma), mouse monoclonal anti-mCherry tag (Novus Biological), rabbit monoclonal anti-mCherry tag (Novus Biological), Alexa Fluor 594-conjugated anti-mouse goat antibody (Invitrogen), Alexa Fluor 488-conjugated anti-rabbit goat antibody (Invitrogen), anti-mouse VisUCyte HRP Polymer (R&D Systems, USA), and HPR-conjugated anti-mouse goat antibody (Invitrogen).