Table 1 Immunohistochemical staining methods
Antibody | Provider | Epitope retrieval | Dilution | Incubation | Signal amplification |
|---|---|---|---|---|---|
HIF-1α, MS-1164, clone 67 (MoAb) | Neomarkers | 15 min in TrisEDTA buffer (pH = 9) at 350 W | 1:100 | 60 min RTa | CSA (DAKO) |
CAIX, NB 100–417 (pAb) | Novus Biologicals | 15 min in citrate buffer (pH = 6) at 350 W | 1:100 | 60 min RT | no |
TNF-α, ab 6671 (pAb) | Abcam | 15 min in TrisEDTA buffer (pH = 9) at 350 W | 1:200 | Overnight 4°C | no |
Apaf-1, RB 9236 (pAb) | Neomarkers | 15 min in citrate buffer (pH = 6) at 350 W | 1:100 | 60 min RT | no |
CD51/αv-integrin, MS 1774, clone CJ00 (MoAb) | Neomarkers | 15 min in EDTA buffer (pH = 8) at 350 W | 1:20 | 60 min RT | no |
CD61/β3-integrin, MS 1842, clone Y2/51 (MoAb) | Neomarkers | 15 min in citrate buffer (pH = 6) at 350 W | 1:25 | Overnight 4°C | no |
CD44/HCAM, MS 668, clone 156-3C11 (MoAb) | Neomarkers | 15 min in citrate buffer (pH = 6) at 350 W | 1:200 | 60 min RT | no |
Osteopontin (MoAb) | Gift from prof. Ann F. Chambers | 15 min in citrate buffer (pH = 6) at 350 W | 1:500 | 60 min RT | No |