TACS® 2 TdT Fluorescein Kit

TUNEL assay cell type - Mouse mammary gland tissue

Experiment
TUNEL assay cell type - Mouse mammary gland tissue
Product
TACS® 2 TdT Fluorescein Kit from Trevigen
Manufacturer
Trevigen

Protocol tips

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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Discussion

Discussion

1 year ago

Author: Ercole Udinesi Italy

Are 10 week old samples in formalin still usable?

I have left my tumor samples in formalin for an extended period of time (around 10 weeks). Do you think I will be able to use them for TUNEL assay and get results? Thank you for your help in advance

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing TUNEL assay cell type - Mouse mammary gland tissue using TACS® 2 TdT Fluorescein Kit from Trevigen.

Paper title
The PARP inhibitors, veliparib and olaparib, are effective chemopreventive agents for delaying mammary tumor development in BRCA1-deficient mice
Full paper
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Manufacturer protocol

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

Download PDF Download manufacturer protocol

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