TABLE 1.
JESS/WES plate loading guide.
| Well | |||
|---|---|---|---|
| One antibody only | w/total protein | Component | Volume |
| A1 | A1 | Biotinylated ladder | 5 μL |
| A2–A25 | A2–A25 | Lens lysate samples | 3 μL |
| B1–B25, C1 | B1, C1–C25, D1 | Antibody diluent | 10 μL |
| B2–B25 | Total protein labeling reagent | 10 μL | |
| C2–C25 | D2–D25 | Primary antibody | 8.5 μL |
| D1 | E1 | Streptavidin-HRP | 10 μL |
| D2–D25 | E2–E25 | Secondary antibody | 10 μL |
| F1–F25 | Total protein streptavidin-HRP | 8 μL | |
| E2–E25 | Row J | Luminol–peroxide | 15 μL (immunoassay) |
| 170 μL (total protein) | |||
| Rows G, H, and I | Row G, H, I | Wash buffer | 500 μL (load after step 10) |
| Row K | RePlex reagent | 300 μL (load after step 10) | |