Anti e cadherin antibody clone hecd 1

In immunohistochemical staining, 10% formalin-fixed and paraffin-embedded tissues were processed into 3-µm-thick sections. The sections were reacted with 1,000-times-diluted anti-S100A16 polyclonal antibody (Abcam, Cambridge, UK) for 2 hours, anti-vimentin antibody (clone V9; DAKO, Glostrup, Denmark) for 2 hours, and 200-times-diluted anti-E-cadherin antibody (clone HECD-1; Takara, Kusatsu, Japan) for 1 hour at room temperature. The details of the procedure were described previously by our laboratory.17 (link)

Immunohistochemistry (IHC) was performed on TMAs containing 4-µm consecutive sections of FFPE gastric tissue samples. After deparaffinization and rehydration, endogenous peroxidases were blocked by incubation in 0.3% hydrogen peroxide solution for 20 min. To expose antigens, the sections were autoclaved in EDTA buffer (pH 9.0) for 10 min and cooled for 30 min. After rinsing in 0.05 M Tris-buffered saline containing 0.1% Tween-20 (pH 7.6), the sections were incubated with an affinity-purified anti-ZEB1 antibody (clone: D80D3; dilution: 1:200; Cell Signaling Technology) and anti-E-cadherin antibody (clone: HECD-1; dilution: 1:1,000; Takara) for 3 h at RT. Samples were then washed three times in PBS and incubated with Dako secondary antibody for 15 min at RT. Horseradish peroxidase-labeled polymer conjugated to a mixture of goat anti-mouse and anti-rabbit immunoglobulin antibodies (Code: K5007; prediluted; Dako, Glostrup, Denmark) was used as the secondary antibody. 3,3-Diaminobenzidine tetrachloride (DAB) was used for color development and sections were counterstained with hematoxylin.