Goat anti human tslpr antibody

Four-week-old female nude mice were purchased from Chinese Academy of Science (Shanghai) and housed in a pathogen-free facility. Experimental procedures were in accordance with the Animal Care and Use Committee at Fudan University. Recombinant human TSLP, rabbit anti-human TSLP antibody and goat anti-human TSLPR antibody applied for immunocytochemistry were purchased from R&D. FITC-conjugated anti-human TSLPR antibody were purchased from eBioscience.

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.