Epr18297

The immunohistochemical technique for the demonstration of caspase-3 expression in the gastric glandular tissues was performed. Initially, alcohol was used to dewax and rehydrate the paraffin-embedded gastric tissue sections. The endogenous peroxidase activity was then inhibited by incubating the sections in 3% hydrogen peroxide. Following that, tissues were incubated with rabbit monoclonal anti-caspase-3 (EPR 18297) (ab 184787) (abcam) and rabbit polyclonal anti-TLR4, 1:50, Santa Cruz, CA, USA. The immune reactivity was visualized using diaminobenzidine (DAB). Based on the percentage of positively stained cells, the immunological response was semi-quantitatively evaluated in 10 randomly selected high-power fields (40×). The results were graded on a scale of 0 to 3, with 0 denoting no staining, 1 denoting positive staining in 30% of cells or HPF, 2 denoting positive staining in 30% to 70% of cells or HPF, and 3 denoting positive staining in more than 70% of cells or HPF.

The primary antibodies, including aFGF (ab169748), NeuN (ab177487), caspase 12 (ab62484), caspase3 (EPR18297), EIF–2α (ab169528), CHOP (ab11419), and GRP78 [EPR4041 (2)] and the secondary antibodies: goat anti-rabbit 488 (ab150077), goat anti-mouse 488 (ab150113), goat anti-rabbit (HRP) (ab6721), and rabbit anti-mouse (HRP) (ab6728) were all bought from Abcam. The DAPI is also provided by Abcam (MC, United Kingdom).