Immunofluorescence of ATG101 in mouse liver tissue with ATG101 Polyclonal Antibody (Product # PA5-21000) at 20 µg/mL.

Western blot was performed using Anti-ATG101 Polyclonal Antibody (Product # PA5-21000) and a 25 kDa band corresponding to Autophagy-related protein 101 was observed across cell lines and tissue. Whole cell extracts (60 µg lysate) of ACHN (Lane 1), MCF7 (Lane 2), Mouse Thymus (Lane 3) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (2 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20000 dilution using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556). Non specific bands were observed at ~20 kDa and ~60 kDa.