Table 1. Protocol for immunohistochemistry.
Antigen-retrieval | Primary antibody | DABd) | |
---|---|---|---|
ZO-1 | 121ºC, 5 min in CBa) | 1:100, 4ºC, over-night | RT, 3 min |
E-cadherin | 121ºC, 5 min in CBa) | 1:150, 4ºC, over-night | RT, 3 min |
Vimentin | 121ºC, 5 min in CBa) | 1:50, 37ºC, 1 hr | RT, 1 min |
N-cadherin | 121ºC, 15 min in TBb) | 1:50, 37ºC, 50 min | RT, 50 sec |
Fibronectrin | RTc), 5 min, in proteinase K | 1:1,000, 37ºC, 50 min | RT, 50 sec |
a) CB, 10 mM citrate buffer, pH6.0, b) TB, 10 mM Tris buffer, pH9.0 with 1 mM EDTA, c) RT, room temperature, d) DAB, 3, 3′-diaminobenzidin/hydrogen peroxide solution.