Table 2. Primary and secondary antibodies used for Western blotting.
CST, Cell Signaling Technology. mAb, monoclonal antibody.
| Primary antibody (clone if mAb) | Vendor | Catalogue # | Lot # | Species | Size (kDa) | Dilution |
|---|---|---|---|---|---|---|
| p110α (C73F8) | CST | 4249 | 7 | rabbit | 110 | 1:1000 |
| pAKT S473 (D9E) | CST | 4060 | 24 | rabbit | 60 | 1:1000 |
| AKT (C67E7) | CST | 4691 | 20 | rabbit | 60 | 1:1000 |
| pPRAS40 T246 (C77D7) | CST | 2997 | 12 | rabbit | 40 | 1:1000 |
| PRAS40 (D23C7) | CST | 2691 | 11 | rabbit | 40 | 1:1000 |
| pS6 S240/S244 (D68F8) | CST | 5364 | 7 | rabbit | 32 | 1:1000 |
| S6 (5G10) | CST | 2217 | 9 | rabbit | 32 | 1:1000 |
| Vinculin (E1E9V) | CST | 13901 | 6 | rabbit | 124 | 1:1000 |
| Secondary antibody | ||||||
| IRDye 800CW Goat anti-Rabbit IgG (H + L) | LI-COR | 926–32211 | D00825-14 | goat | 1:20,000 |