Table 2. Antibodies and immunoreagents.
| Antibody | Dilution | Manufacturer |
|---|---|---|
|
Primary antibodies | ||
| Anti-mouse ephrin-A1 (goat, poly) |
1:2,500** |
R&D Systems |
| Anti-human ephrin-A4 (goat, poly) |
1:2,500** |
R&D Systems |
| Anti Paired Box-2 (Pax2; rabbit, poly)* |
1:100 |
Covance |
| Control antibody |
||
| Normal goat IgG |
1:25,000** |
R&D Systems |
|
Secondary antibodies | ||
| Biotinylated horse anti-goat (0.3% PBSR) |
1:200 |
Zymed |
| Biotinylated goat anti-rabbit (0.3% PBSR) |
1:200 |
Zymed |
|
Visualization | ||
| Conjugated goat anti-rabbit Alexa 594 (0.3% PBSR) |
1:1,000 |
Invitrogen |
| Streptavidin Alexa 488 | 1:1,000 | Invitrogen |
Antibodies raised against the two ligands to Eph-A6, ephrin-A1, and -A4, were used to localize these proteins in sections of fetal macaque retina. Anti-Pax2 double immunolabel was used to localize immature astrocytes, while background labeling was visualized using normal goat IgG antibody. *Pax2 is cross-reactive to human and other species. **IgG concentration applied to tissue is equal to ephrin antibodies given the reconstituted antibodies stocks were 0.1 mg/ml for ephrins and 1 mg/ml for IgG.