TACS® 2 TdT Fluorescein Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add Proteinase K Solution and incubate 15 to 30 minutes at room temperature, Add Labeling Reaction Mix and incubate at 37˚C for 1 hour in a humidity chamber.	If no staining in TACS-Nuclease, increase incubation time with Proteinase K (up to 60 minutes) or Cytonin™ (up to overnight at 4 oC)

Protocol tips
Add Proteinase K Solution and incubate 15 to 30 minutes at room temperature, Add Labeling Reaction Mix and incubate at 37˚C for 1 hour in a humidity chamber.
Downstream tips
If no staining in TACS-Nuclease, increase incubation time with Proteinase K (up to 60 minutes) or Cytonin™ (up to overnight at 4 oC)

Publication protocol

For biomarker studies, mice were fed olaparib (200 mg/kg) intermittently in diet, as described above, and sacrificed at 18, 24, or 30 weeks of age. Mammary glands were harvested for immunohistochemistry. To measure proliferation, mice were injected intraperitoneally with bromodeoxyuridine (BrdU; BD Pharmingen) in sterile saline (1 mg/mouse) two hours before sacrifice. Tissues were then fixed in 10% phosphate-buffered formalin for at least 48 hours before they were embedded in paraffin blocks and sectioned. BrdU-positive cells were stained using the BrdU in situ detection kit (BD Pharmingen). To identify cells undergoing apoptosis, TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling staining) was performed on tissue sections using the TdT2-TACS in situ apoptosis detection kit (Trevigen). Six mice per group were used in the 24-week biomarker studies while five mice per group were used in the 30-week biomarker studies. To quantitate the number of BrdU-positive cells, 11–26 hyperplastic areas as well as 56–130 ductal areas from each group were counted. To assess apoptosis, similar numbers of hyperplastic areas and ducts were evaluated. PARP activity in mammary glands from these same mice was also measured using the HT Universal Colorimetric PARP Assay Kit with Histone-coated Strip Wells (Trevigen).

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Manufacturer protocol

Download the product protocol from Trevigen for TACS® 2 TdT Fluorescein Kit below.

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