TABLE 2.
Summary of antibodies and conditions for immunohistochemistry of drug transporters in human ocular tissues
| Transporter | Primary Antibody | Dilution | Incubation Condition | Secondary Antibody | Antigen Retrieval Procedure |
|---|---|---|---|---|---|
| PEPT-1 | Anti-human goat PEPT-1 Ab (Santa Cruz Biotech, Santa Cruz, CA) | 1:200 | 60 min at 37°C | Poly-AP anti-goat IgG | High pH heat-induced antigen retrieval (10 mM Tris-HCl, 1mM EDTA, pH 9.0 at 95°C for 20 min) |
| PEPT-2 | Anti-human rabbit PEPT-2 Ab (Santa Cruz Biotech) | 1:200 | Overnight at 4°C | Poly-AP anti-rabbit IgG | Low pH heat-induced antigen retrieval (10 mM citrate buffer, pH 6.0 at 95°C for 20 min) |
| OCT-1 | Anti-human rabbit OCT-1 Ab (Santa Cruz Biotech) | 1:200 | Overnight at 4°C | Poly-AP anti-rabbit IgG | High pH heat-induced antigen retrieval (10 mM Tris-HCl, 1mM EDTA, pH 9.0 at 95°C for 20 min) |
| OCT-2 | Anti-human rabbit OCT-2 Ab (Santa Cruz Biotech) | 1:200 | 60 min at 37°C | Poly-AP anti-rabbit IgG | High pH heat-induced antigen retrieval (10 mM Tris-HCl, 1 mM EDTA, pH 9.0 at 95°C for 20 min) |
| ATB0,+ | Anti-human rabbit ATB0,+ Ab (Medical and Biologic Laboratories, Nagoya, Japan) | 1:5000 | 60 min at 37°C | Poly-AP anti-rabbit IgG | High pH heat-induced antigen retrieval (10 mM Tris-HCl, 1 mM EDTA, pH 9.0 at 95°C for 20 min) |
| MCT-1 | Anti-human goat MCT1 Ab (Santa Cruz Biotech) | 1:200 | 60 min at 37°C | Poly-AP anti-goat IgG | Low pH heat-induced antigen retrieval (10 mM citrate buffer, pH 6.0 at 95°C for 20 min) |
| MCT-3 | Anti-human rabbit MCT3 Ab (Santa Cruz Biotech) | 1:200 | 60 min at 37°C | Poly-AP anti-rabbit IgG | No antigen retrieval |
Ab, antibody; ATB0,+, neutral and basic amino acid transporter; MCT, monocarboxylate transporter; OCT, organic cation transporter; PEPT, peptide transporter.