Table 2.
Summary of antibody dilutions and conditions used in the immunoblotting analysis of PHF-Tau (AT8), total tau, DYRK1A, MAP2, GSK3β, cdk5 and mitochondrial biogenesis and synaptic proteins in HT22 cells, HT22 cells treated with phosphorylated tau ligand 1, cells transfected with cDNA mutant Tau, mutant Tau cDNA transfected HT22 cells treated with phosphorylated tau ligand 1
| Marker | Primary antibody— species and dilution | Purchased from company, city & state | Secondary antibody, dilution | Purchased from company, city & state |
|---|---|---|---|---|
| DYRK1A | Rabbit Polyclonal 1:500 | ABCAM, Cambridge, MA 02139–1517, USA | Goat 1:10 000 anti-rabbit | ABCAM, Cambridge, MA 02139–1517, USA |
| Total tau | Mouse Monoclonal 1:200 | Thermo Fisher Scientific, Waltham, MA | Goat anti-mouse, HRP 1:60 000 | Thermo Fisher Scientific, Waltham, MA |
| Tau-pSer202/ Thr205 (AT8) | Mouse Monoclonal 1:500 | Thermo Fisher Scientific, Waltham, MA | Donkey anti-mouse 1:10 000 | Thermo Fisher Scientific, Waltham, MA |
| MAP2 | Mouse Monoclonal Antibody 1:500 | Thermo Fisher Scientific, Waltham, MA | Goat anti-mouse, HRP 1:4000 | Thermo Fisher Scientific, Waltham, MA |
| GSK-3β (D5C5Z) | Rabbit Monoclonal Antibody 1:1000 | Cell Signaling Technology, MA 01923 | Donkey anti-rabbit HRP 1:10 000 | Cell Signaling Technology, MA 01923 |
| CDK5 | Rabbit Polyclonal Antibody 1:1000 | Cell Signaling Technology, MA 01923 | Goat 1:10 000 anti-rabbit | Cell Signaling Technology, MA 01923 |
| PGC1α | Rabbit Polyclonal 1:500 | Novus Biological, Littleton, CO | Donkey anti-rabbit HRP 1:10 000 | GE Healthcare Amersham, Piscataway, NJ |
| Nrf1 | Rabbit Polyclonal 1:300 | Novus Biological, Littleton, CO | Donkey anti-rabbit HRP 1:10 000 | GE Healthcare Amersham, Piscataway, NJ |
| Nrf2 | Rabbit Polyclonal 1:300 | Novus Biological, Littleton, CO | Donkey anti-rabbit HRP 1:10 000 | GE Healthcare Amersham, Piscataway, NJ |
| TFAM | Rabbit Polyclonal 1:300 | Novus Biological, Littleton, CO | Donkey anti-rabbit HRP 1:10 000 | GE Healthcare Amersham, Piscataway, NJ |
| Mfn1 | Rabbit Polyclonal 1:300 | Novus Biological, Littleton, CO | Donkey anti-rabbit HRP 1:10 000 | GE Healthcare Amersham, Piscataway, NJ |
| Mfn2 | Rabbit Polyclonal 1:300 | Novus Biological, Littleton, CO | Donkey anti-rabbit HRP 1:10 000 | GE Healthcare Amersham, Piscataway, NJ |
| OPA1 | Rabbit Polyclonal 1:300 | Novus Biological, Littleton, CO | Donkey anti-rabbit HRP 1:10 000 | GE Healthcare Amersham, Piscataway, NJ |
| DRP1 | Rabbit Polyclonal 1:300 | Novus Biological, Littleton, CO | Donkey anti-rabbit HRP 1:10 000 | GE Healthcare Amersham, Piscataway, NJ |
| FIS1 | Rabbit Polyclonal 1:300 | Novus Biological, Littleton, CO | Donkey anti-rabbit HRP 1:10 000 | GE Healthcare Amersham, Piscataway, NJ |
| PSD95 | Rabbit Monoclonal 1:300 | Novus Biological, Littleton, CO | Donkey anti-rabbit HRP 1:10 000 | GE Healthcare Amersham, Piscataway, NJ |
| SYN | Rabbit Monoclonal 1:400 | Novus Biological, Littleton, CO | Donkey anti-rabbit HRP 1:10 000 | GE Healthcare Amersham, Piscataway, NJ |
| B-Actin | Mouse Monoclonal 1:500 | Sigma-Aldrich, St Luis, MO | Sheep anti-mouse HRP 1:10 000 | GE Healthcare Amersham, Piscataway, NJ |