Immunoblotting conditions for the detection of intestinal P450 isoforms
P450 | 1A1 | 2A6 | 2B6 | 2C | 2D6 | 2E1 | 2J2 | 3A4 | 3A5 |
|---|---|---|---|---|---|---|---|---|---|
| 1° Antibody | |||||||||
| Host | Goat | Mouse | Rabbit | Rabbit | Mouse | Goat | Rabbit | Rabbit | Rabbit |
| Dilution | 1:1000 | 1:1000 | 1:1000 | 1:1000 | 1:3000 | 1:1500 | 1:1000 | 1:500 | 1:3000 |
| Incubation time (h) | 3 | 3 | 1 | 2 | 1 | 3 | 1 | 1 | 2 |
| 2° Antibody | |||||||||
| Dilution | 1:2000 | 1:2000 | 1:1000 | 1:1000 | 1:2000 | 1:2000 | 1:2000 | 1:500 | 1:3000 |
| Incubation time (h) | 2 | 3 | 1 | 2 | 1 | 3 | 1 | 1 | 2 |
| Mass of HIM loaded/well (μg) | 40 | 40 | 40 | 40 | 40 | 40 | 40 | 5 | 10 |