Anti chmp4b

Rabbit anti-Hrs was raised using GST–Hrs as an immunogen and was generated by Eurogentec, Southampton, UK. The antibody was affinity-purified against GST–Hrs and adsorbed against GST. Sheep anti-VPS28 was as described previously (Bishop et al., 2002 (link)). Mouse anti-Hrs was from Enzo. Rabbit anti-EEA1 was from Cell Signaling Technology. Mouse anti-EEA1 was from BD Biosciences. Mouse anti-EGFR was Mab 108, purified from supernatants of the hybridoma clone HB-9764 (ATCC; Washington DC), grown in RPMI medium and 10% FCS under 5% CO₂, and antibody affinity-purified using Protein-G–Sepharose. Rabbit anti-APPL1 and anti-CHMP4B were from Proteintech Europe (Manchester, UK). Rabbit anti-clathrin was from Abcam (Cambridge, UK). Fluorescent EGF was from Invitrogen (Paisley, Scotland) and secondary antibodies were from Jackson ImmunoResearch (West Grove, PA). Human SNX15 in pEGFP-C1 (Clontech) was a gift from Chris Danson and Pete Cullen, University of Bristol.

Antibodies used were: rabbit polyclonal anti-TMEM16F (Sigma-Aldrich, HPA038958; IB 1:1000); mouse monoclonal anti-SMS2 [7D10] (Santa Cruz, sc-293384; IB 1:1000); mouse monoclonal anti-nSMase-2 (Santa Cruz, sc-166637; IB: 1:1000); mouse monoclonal anti-LAMP1 [H4A3] (Santa Cruz, sc-20011; IF 1:200); rabbit polyclonal anti-CHMP4B (Proteintech, 13683-1-AP; IF 1:300); mouse monoclonal anti-ALIX [3A9] (Biolegend, 634501, IB 1:1000); mouse monoclonal anti-Actin (Sigma-Aldrich, A1978; IF 1:1200; IB 1:10,000); rabbit monoclonal anti-Na/K-ATPase [EP1845Y] (Abcam, ab-76020; IF 1:600); mouse monoclonal anti-TSG101 [C-2] (Santa Cruz, sc-7964; IB 1:1000); mouse monoclonal anti-V5 [R960-25] (Invitrogen, r96025; IF 1:400; IB 1:1000); rabbit polyclonal anti-GFP (Novus Biologicals, NB600-303; IF 1:250); HRP-conjugated goat anti-mouse IgG (Thermo Fisher Scientific, 31430; IB 1:5000); HRP-conjugated goat anti-rabbit IgG (Thermo Fisher Scientific; 31460; IB 1:5000); Cyanine Cy™2-conjugated donkey anti-mouse IgG (Jackson ImmunoResearch Laboratories, 715-225-150; IF 1:400); Cyanine Cy™2-conjugated donkey anti-rabbit IgG (Jackson ImmunoResearch Laboratories, 711-225-152; IF 1:400); Cyanine Cy™3-conjugated donkey anti-rabbit IgG (Jackson ImmunoResearch Laboratories, 715-165-152; IF 1:400); and Cyanine Cy™3-conjugated donkey anti-mouse IgG (Jackson ImmunoResearch Laboratories, 715-165-150; IF 1:400).