Data and conditions of primary and secondary antibodies used for Western blotting
Anti-pRb and p21 were obtained from BD Biosciences Transduction Laboratories (Lexington, KY). Anti-p-4E-BP1 (Thr37/46) was obtained from Cell Signaling Technology Inc. (Beverly, MA). The remaining primary antibodies were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA). All secondary antibodies were obtained from Pierce (Rockford, IL).
Primary Antibody Anti- | SDS-PAGE Conditions | Primary Antibody Dilution | Secondary Antibody | Secondary Antibody Dilution |
|---|---|---|---|---|
| % | ||||
| pRb | 7.5 | 1:2500 | Goat-anti-mouse | 1:15,000 |
| p27 | 15 | 1:1000 | Donkey-anti-rabbit | 1:64,000 |
| p70s6k | 10 | 1:100 | Rabbit-anti-goat | 1:50,000 |
| p-p70s6k (Ser411) | 10 | 1:100 | Goat-anti-mouse | 1:15,000 |
| 4E-BP1 | 15 | 1:2000 | Rabbit-anti-goat | 1:50,000 |
| p-4E-BP1 (Ser65/Thr70) | 15 | 1:200 | Donkey-anti-rabbit | 1:64,000 |
| p-4E-BP1 (Thr37/46) | 15 | 1:200 | Donkey-anti-rabbit | 1:64,000 |
| p21 | 12 | 1:200 | Goat-anti-mouse | 1:7500 |