Key Resources Tables:
| Antibodies | ||
|---|---|---|
| Name | Product ID | Dilution |
| Anti-Alpha-synuclein antibody (MJFR1) | Abcam (ab138501) | IF: 1:300 – 1:1000* WB: 1:1000 |
| Anti-Synapsin-1 | SySy (106-011) | IF: 1:500 WB: 1:1000 |
| Anti-α-synuclein Antibody (syn211) | Santa Cruz (sc-12767) | WB: 1:200 |
| Anti-α-synuclein (phospho S129) Antibody (81a) | Provided by Center for Neurodegenerative Disease Research (CNDR) at University of Pennsylvania | IF: 1:5000 |
| Anti-Human Osteoactivin/GPNMB Antibody | RnD Systems (AF2550) | IF: 1:500 IHC: 1:200 WB: 1:1000 |
| Anti-GPNMB (E1Y7J) Rabbit mAb | Cell Signaling Technology (#13251) | WB: 1:1000 |
| Anti-GPNMB Antibody (D-9) | Santa Cruz (sc-271415) | IF: 1:500 WB: 1:250 |
| Anti-beta Actin antibody (AC-15) | Abcam (ab6276) | WB: 1:5000 |
| Anti-beta-III Tubulin Antibody | Novus Biologicals (NB100-1612) | IF: 1:500 |
| Anti-Flag M2 Mouse Monoclonal Antibody | Sigma Aldrich (F1804) | IF: 1:1000 |
| Anti-GAPDH Antibody | Advanced Immunochemical Anti-GAPDH | WB: 1:3000 |
| Anti-LAMP1 Antibody (H4A3) | University of Iowa Hybridoma Bank | IF: 1ug/mL |
| Goat Anti-Chicken Alexa Fluor 647 | Invitrogen (A-32933) | IF: 1:1000 |
| Goat Anti-Rabbit Alexa Fluor 488 | Invitrogen (A-21206) | IF: 1:1000 |
| Donkey Anti-Mouse Alexa Fluor 594 | Invitrogen (A-21203) | IF: 1:1000 |
| DRAQ5 | Thermo-Fischer (62251) | IF: 1:5000 |
| Goat Anti-Mouse HRP | Jackson Immuno Research (115-035-062) | WB: 1:3000 |
| Goat Anti-Rabbit HRP | Jackson Immuno Research (111-035-144) | WB: 1:3000 |
| Biotinylated Rabbit Anti-Goat | Vector Laboratories (BA-5000-1.5) | IHC: 1:5000 |
| Cell Lines | |||
|---|---|---|---|
| Name | Species | Obtained from | Citations |
| HeLa | Homo sapiens | Michael S. Marks | |
| QBI293 | Homo sapiens | Penn Center for Neurodegenerative Disease Research (CNDR) | |
| iPSC-N | Homo sapiens | George Church | (22) |
| Deglycosylation and dephosphorylation | ||
|---|---|---|
| Name | Product ID | Protocol |
| PNGase F (Deglycosylation) | New England Biolabs Inc., Catalog # P0704L | As per product instructions |
| Lambda Protein Phosphatase (Dephosphorylation) | New England Biolabs Inc., Catalog #P0753S | As per product instructions |
*
MJFR1 was used at 1:300 to stain endogenous α-synuclein in iPSC-N and 1:1000 to stain overexpressed α-synuclein in HEK293 and HeLa cells.