Texasred conjugated secondary antibody

For immunofluorescence study, the gastric tissue samples were fixed in 4% paraformaldehyde solution for 48 h, dehydrated in ascending alcohol series. It was embedded in paraffin wax and sectioned at 5 μm thickness using a microtome. The tissue sections were deparaffinized with xylene followed by rehydration with descending alcohol series. Antigen retrieval was done by trypsin (0.05% trypsin, 0.1% CaCl₂), and blocking was performed using 5% BSA in TBS (20 mM Tris-HCl, pH 7.4 containing 150 mM NaCl) for 2 h at room temperature followed by the incubation overnight at 4°C in primary antibody solution (1:200 dilutions in TBS with 1% BSA) in a humid chamber. The tissue sections were washed four times with TBST (20 mM Tris-HCl, pH 7.4 containing 150 mM NaCl and 0.025% Triton X-100) followed by incubation with Texas red-conjugated secondary antibody (Santa Cruz Biotechnology, USA) solution (1:400 dilutions in TBS containing 1% BSA) for 2 h at room temperature. The sections were counter-stained using DAPI and images were observed in an Olympus microscope. Images at 20× and 60× magnification were captured using Camedia software (E− 20P 5.0 Megapixel) and processed under Adobe Photoshop version 7.0.