Brdu red dna fragmentation assay kit

Paraffin-embedded tumor tissues were sectioned to a thickness of 5 μm,
deparaffinized with xylene, rehydrated through xylene and graded alcohol and
blocked with 5% bovine serum albumin. Immunofluorescence (IF) staining was
conducted with the indicated antibodies and fluorochrome-conjugated secondary
antibodies (Alexa-488 and 564). Apoptotic cells were identified by terminal
deoxynucleotidyl transfer-mediated dUTP nick-end labeling (TUNEL) staining using
an in situ BrdU-Red DNA fragmentation assay kit (Abcam,
Cambridge, UK). Nuclei were counterstained with DAPI. The slides were examined
in a blinded manner and randomly chosen fields were photographed at 400x
magnification. The immune-positive cells were quantified with a Carl Zeiss
AxioImager microscope and Image M1 Software (Carl Zeiss, Jena, Germany).

Sixteen weeks after curdlan injection (experimental end-point), eyeball tissues from control and treated mice were fixed in 10% buffered formalin and paraffin-embedded. Sections (4 μm in thickness) were cut and stained with hematoxylin and eosin. For detection of apoptosis and necrotic cell death in eyeball tissues, in situ BrdU-Red DNA Fragmentation assay kit (abcam) was used according to the manufacturer's instructions. Stained slides were scanned with the multispectral Vectra scanner (Perkin Elmer).