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. 2014 Jun 16;76(10):1321–1327. doi: 10.1292/jvms.14-0104

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.