Clone mf1

Three monoclonal antibodies (MAbs) that recognised human tetraspanins CD9 (clone 602.29), CD63 (clone H5C6), CD81 (clone 1D6, Bio-Rad, USA), and IgG1 isotype control (clone JC1) were used to treat A549 cells. MAbs that recognised mouse tetraspanins, including MAbs specific to CD9 (clone MF1, Bio-Rad), CD63 (clone NVG-2, BioLegend Inc., San Diego, CA, USA), CD81 (clone Eat2, Bio-Rad), and rat IgG2b, rat IgG2a, and hamster IgG1 isotype controls were used to treat J774A.1 cells. The production of recombinant glutathione S-transferase (GST) fusion proteins with CD9, CD63, and CD81 tetraspanin EC2 extracellular domains were previously described^{30 (link)}.

Monoclonal antibodies (mAb) against mouse tetraspanins were CD9 (Cat. No. MCA2749, clone MF1 Bio-Rad), CD63 (Cat. No. 143902, clone NVG-2, Biolegend), CD81 (Cat. No. MCA1846, clone Eat2 Bio-Rad), and matching isotype controls rat IgG2b (Bio-Rad), rat IgG2aκ (Biolegend) and hamster IgG1 (Bio-Rad), respectively. MAb to other cell surface molecules were CD36 (Cat. No. 102602, clone HM36), CD44 (Cat. No. 103002, clone IM7) CD47 (Cat. No. 127502, clone miap301) CD98 (Cat. No. 128202, clone RL388), CD172a (Cat. No. 144002, clone P84) (all from Biolegend), and DC-STAMP (Cat. No. MABF39, clone 1A2 Millipore). Isotype controls were Armenian hamster IgG, rat IgG2bκ, rat IgG2aκ, rat IgG2aκ, rat IgG1κ, (Biolegend), and IgG2aκ (Millipore), respectively. The secondary antibodies used for flow cytometry were anti-rat IgG-FITC (Cat. No. F-9387, Sigma) and anti-hamster IgG-FITC (Cat. No. MCA2357, Bio-Rad). All antibodies were used at saturating binding concentrations.