Tunel in situ cell death detection kit pod

For the detection of TSLPR expression by colon cancer cells, the coverslips were put into 6-well plate before the seeding of colon cancer cells, the cell climbing slices were taken out and fixed in 4% paraformaldehyde. Paraffin-embedded human colon sections and climbing slices were incubated with rabbit anti-human TSLP antibody and goat anti-human TSLPR antibody (1:100 dilution, R&D), respectively. The secondary antibody used was HRP-conjugated goat anti-rabbit IgG and mouse anti-goat IgG (1:200 dilution, ebioscience), respectively. Slices added with PBS instead of the primary antibodies were used as negative controls. The intensity of TSLP staining was evaluated in a blind manner by two independent investigators and graded by a 5-scale system (0, no signal; 1, weak; 2, moderate; 3, strong; 4, very strong; and 5, extremely strong). Paraffin-embedded sections of tumor excised from the mice were stained with Hematoxylin & Eosin. The necrotic areas were examined under microscope. Detection of fluorescence binding to 3′-OH of DNA fragments was performed with the TUNEL in situ cell death detection kit-POD (Roche), according to the manufacturer's instruction.

Nuclei with DNA-fragmentation were stained with TUNEL In Situ Cell Death Detection Kit (POD, Roche, Mannheim, Germany) according to the manufacturer's protocol. The fluorescence signal was converted and then stained with diaminobenzidine (AXXORA) and slides were counterstained with Quick hemalaun kit (Vector) [19 (link)].