Table 4.
Antibodies used in this work.
| Primary antibodies | |||||
|---|---|---|---|---|---|
| Antigen | Species | Manufacturer | Identifier | Dilution | Application |
| FLAG | Mouse | Sigma-Aldrich | F1804 | 1:5,000 | WB |
| HA | Rabbit | CST | 3724 | 1:1,000 | WB |
| NEK1 | Rabbit | ProteinTech | 27146-1-AP | 1:1,000 | WB |
| NEK1 (aa 700–1,286) | Sheep | MRC-PPU | SA353 | 1:2,000 | WB |
| NEK1 (aa 900–1,286) | Sheep | MRC-PPU | SA354 | 1:2,000 | WB |
| NEK1 | Rabbit | Bethyl Laboratories | A304-570A | 1:2,000 | WB |
| C21ORF2 (full length) | Sheep | MRC-PPU | DA066 | 0.1 μg/ml | WB |
| C21ORF2 (full length) | Rabbit | ProteinTech | 27609-1-AP | 1:1,000 | WB |
| GAPDH | Rabbit | CST | 2118 | 1:10,000 | WB |
| α-Tubulin | Mouse | CST | 3873 | 1:10,000 | WB |
| GFP | Chicken | Abcam | Ab13970 | 1:1,000 | WB |
| HIF-1α | Mouse | BD | 610959 | 1:1,000 | WB |
| ATG14 | Rabbit | CST | 96752 | 1:1,000 | WB |
| ATG14 pSer29 | Rabbit | CST | 92340 | 1:1,000 | WB |
| LC3A/B | Rabbit | CST | 4108 | 1:1,000 | WB |
| FLAG | Mouse | Sigma-Aldrich | F1804 | 1 μg/1 mg lysate | IP |
| Mouse IgG1 (isotype control for FLAG antibody) | Mouse | CST | 5415 | 1 μg/1 mg lysate | IP |
| HA beads | Mouse | MRC-PPU | Frankenbody | 15 μl 50% slurry/ 1 mg lysate | IP |
| Mouse IgG2b (isotype control for HA beads) | Mouse | CST | 53484 | 1 μg/1 mg lysate | IP |
| NEK1 | Rabbit | ProteinTech | 27146-1-AP | 2 μg/1 mg lysate | IP |
| NEK1 (aa 900–1,286 aa) | Sheep third bleed | MRC-PPU | SA354 | 1 μg/1 ml lysate | IP |
| NEK1 | Rabbit | Bethyl Laboratories | A304-570A | 2 μg/1 mg lysate | IP |
| C21ORF2 (full length) | Sheep fourth bleed | MRC PPU | DA066 | 1 μg/1 mg lysate | IP |
| C21ORF2 (full length) | Rabbit | ProteinTech | 27609-1-AP | 2 μg/1 mg lysate | IP |
| Rabbit IgG | Rabbit | CST | 2729 | 2 μg/1 mg lysate | IP |
| Sheep IgG | Sheep | Abcam | ab37385 | 1 μg/mg lysate | IP |
| NEK1 (aa 900–1,286) | Sheep | MRC PPU | SA344 | 1:100 | IF |
| C21ORF2 (full length) | Sheep | MRC PPU | DA066 | 1:200 | IF |
| GTU88 (γ-tubulin) | Mouse | Sigma-Aldrich | T6557 | 1:500 | IF |
| ɣH2AX | Mouse | Merck Millipore | 05-636 | 1:2,000 | IF |
| ARL13B | Rabbit | ProteinTech | 17711-1-AP | 1:200 | IF |
| Pericentrin | Mouse | Abcam | ab28144 | 1:500 | IF |
| Secondary antibodies | |||||
| Antigen | Species | Manufacturer | Code | Dilution | Application |
| Anti-mouse 680 | Donkey | LI-COR | 926-68072 | 1:10,000 | WB |
| Anti-mouse 800 | Donkey | LI-COR | 926-32212 | 1:10,000 | WB |
| Anti-rabbit 680 | Donkey | LI-COR | 926-68073 | 1:10,000 | WB |
| Anti-rabbit 800 | Donkey | LI-COR | 926-32213 | 1:10,000 | WB |
| Anti-sheep AF680 | Donkey | Thermo Fisher Scientific | A21102 | 1:10,000 | WB |
| Anti-sheep Dylight 800 | Donkey | Rockland | 613-745-168 | 1:10,000 | WB |
| Anti-chicken 800 | Donkey | LI-COR | 926-32218 | 1:10,000 | WB |
| Protein G Dylight 800 | n/a | Rockland | PG00-45 | 1:10,000 | WB |
| Anti-mouse AF488 | Donkey | Thermo Fisher Scientific | A21202 | 1:1,000 | IF |
| Anti-mouse AF647 | Donkey | Thermo Fisher Scientific | A32787 | 1:1,000 | IF |
| Anti-mouse AF647 | Goat | Thermo Fisher Scientific | #A21235 | 1:500 | IF |
| Anti-rabbit AF488 | Donkey | Thermo Fisher Scientific | A21206 | 1:1,000 | IF |
| Anti-rabbit AF647 | Donkey | Thermo Fisher Scientific | A31573 | 1:1,000 | IF |
| Anti-sheep AF488 | Donkey | Thermo Fisher Scientific | A11015 | 1:500 | IF |