Goat anti biotin horseradish peroxidase hrp conjugated antibody

Proteins were resolved by SDS-PAGE in 4–12% Bolt Bis-Tris Plus pre-cast gels (Invitrogen, Carlsbad, CA, USA) and then transferred to PVDF membranes using the iBlot system (Invitrogen). Membranes were developed using the Clarity Western ECL substrate kit (Bio-Rad, Hercules, CA, USA) and the ChemiDocTM MP imaging system (Bio-Rad, Hercules, CA, USA). The following primary antibodies were used: goat anti biotin-horseradish peroxidase (HRP)-conjugated antibody (Cell Signaling Technology, Danvers, MA, USA; catalogue number 7075) at 1:1000; USP1 (Cell Signaling Technology, Danvers, MA, USA; catalogue number 8033); rabbit polyclonal anti-USP7 antibody (Bethyl laboratories, Montgomery, TX, USA; catalogue number A300-033A) at 1:3000; rabbit polyclonal anti-USP9X antibody (ProteinTech, Rosemont, IL, USA; catalogue number 55054-1-AP) at 1:1000; rabbit monoclonal anti-USP11 (Abcam, Cambridge, UK; catalogue number ab109232); mouse monoclonal anti-Tubulin beta antibody (Developmental Studies Hybridoma Bank, Iowa City, IA, USA; catalogue number E7) at 1:1000). The following secondary antibodies were used: goat anti-mouse-HRP-labelled antibody (Invitrogen, Carlsbad, CA, USA; catalogue number 62-6520) at 1:4000; goat anti-rabbit-HRP labelled antibody (Cell Signaling Technology, Danvers, MA, USA; catalogue number 7074).

The following antibodies were used against Drosophila proteins: goat anti-biotin-horseradish peroxidase (HRP) conjugated antibody (Cell Signalling); chicken polyclonal anti-BirA antibody (Sigma); rabbit polyclonal anti-Parkin antibody [44 (link)]; mouse monoclonal anti-Syx1A antibody (DSHB); rabbit polyclonal anti-RdhB [45 (link)]; rabbit polyclonal anti-ArgK [46 (link)]; rabbit polyclonal anti-Vps4 [47 (link)]; rabbit polyclonal anti-Fax antibody (a gift from Eric Liebl); rabbit polyclonal anti-Ubiquitin antibody (Sigma). The following antibodies were used against Human proteins: goat polyclonal anti-VPS35 antibody (Abcam); mouse monoclonal anti-Cleaved Parp-85 fragment (Cell Signaling); mouse monoclonal anti-Parkin (Santa Cruz); rabbit polyclonal anti-PINK1 (Novus Biologicals); rabbit polyclonal anti-Miro1 (Sigma); rabbit polyclonal anti-Tim44 (Sigma); rabbit polyclonal anti-Tom20 (Sigma); mouse monoclonal (Abcam) and rabbit polyclonal (Sigma) anti-Actin. For monitoring the GFP pull-downs the following antibodies were used: monoclonal mouse anti-GFP antibody (Roche) and monoclonal mouse anti-Flag M2-HRP conjugated antibody (Sigma). Anti-mouse, rabbit and chicken HRP labelled secondary antibodies (Jackson ImmunoResearch Laboratories) and anti-guinea pig (Invitrogen) were used; and anti-mouse, rabbit and sheep IR 680 and IR800-coupled antibodies (LI-COR Biosciences).