Table 1.
Lysosomal network proteins screened, their most frequent cellular compartment association and the primary antibodies used for Western blot. All antibodies were used at a 1:1,000 dilution unless stated otherwise
| Target | Product number | Company | Address | Compartment association |
|---|---|---|---|---|
| Atg4B | Code OSA00033 W/ID tag RB270-270108-WS | Osenses | Keswick, SA, Australia | Autophagosomes |
| Atg9 | Code OSA00042 W/ID tag Rb273-270108-WS | Sigma-Aldrich | St. Louis, MO, USA | Autophagosomes |
| Atg5 | A0731 | Autophagosomes | ||
| EEA1 | E4156 | Endosomes | ||
| Rab5a | R4654 | Endosomes | ||
| Atg7 | ab53255 | Abcam | Cambridge, UK | Autophagosomes |
| IDE | ab133561 | Cytosolic | ||
| Rab9 | ab2810 | Endosomes | ||
| MHC class IIa | ab55152 | Lysosomes | ||
| Atg6a | sc11427 | Santa Cruz Biotechnology | Santa Cruz, CA, USA | Autophagosomes |
| hsc70 | sc-24 | Lysosomes | ||
| DRAM | sc-81713 | Lysosomes | ||
| LIMP-2 | sc-55571 | Lysosomes | ||
| Rab4 | sc-312 | Endosomes | ||
| Rab7 | sc-10767 | Endosomes | ||
| TFEB | sc-4878 | Cytosolic/Nuclear | ||
| V-ATPase | sc-55544 | Lysosomes | ||
| PICALM | NBP1-86658 | Novus Biologicals | Littleton, CO, USA | Endosomes |
| LC3 | NB600-1384 | Autophagosomes | ||
| NPC1 | NB400-148 | Lysosomes | ||
| Cathepsin A | AF1049 | R&D Systems Inc. | Minneapolis, MN, USA | Lysosomes |
| Cathepsin B | 01-12-030102 | Athens Research and Technology | Athens, GA, USA | Lysosomes |
| Cathepsin D | 01-12-030104 | Lysosomes | ||
| Cathepsin L | 01-12-030112 | Lysosomes | ||
| CD63 | 556019 | BD Biosciences | Franklin Lakes, NJ, USA | Lysosomes |
| Flotillin 1 | 610821 | Endosomes | ||
| LAMP-1 | 9835-01 | Southern Biotech | Birmingham, AL, USA | Lysosomes |
| LAMP-2 | 9840-01 | Lysosomes | ||
| Proteasome 20S | BML-PW8105 | Enzo Life Sciences Inc. | Farmingdale, NY, USA | Cytosolic |
| Rab3 | 107011 | Synaptic Systems GmbH | Goettingen, Germany | Endosomes |
| SYT7 | 105172 | Lysosomes | ||
| Tsg 101 | GTX70255 | GeneTex Inc. | San Antonio, TX, USA | Endosomes |
| M6PR | 10R-M105a | Fitzgerald | North Acton, MA, USA | Endosomes |
| ASMase | 3687 | Cell Signaling Technology | Boston, MA, USA | Lysosomes |
aUsed at 1:500 dilution