Antibody Preparation and Lysosome Staining

Rabbit anti-cystatin F polyclonal antibody, recombinant cistatin F and mouse 1D10 anti-cathepsin H Mab was prepared by our group in Ljubljana as reported [45 (link), 50 (link)]. Rabbit N1 antibody against N-terminal of cystatin and Δ15N cystatin F were prepared at Dundee University, Dundee, UK, [54 (link)]. Goat anti-cathepsin C pAb was from R&D Systems (Minneapolis, MN), mouse anti-GAPDH mAb was from Invitrogen and rabbit anti-CRK-L pAb was from Sigma. The dye identifying the lysosomes, LysoTracker, was from Molecular Probes (Eugene, OR). Secondary labelled antibodies goat anti-rabbit Alexa Fluor 488 and donkey anti-goat Alexa Fluor 555 were from Molecular Probes (Eugene, OR). Antibodies to CD16 were purchased from Biolegend (San Diego, CA). FITC anti-human/mouse granzyme B antibody was purchased from Biolegend (San Diego, CA).