TUNEL Assay_Molecular Biology Services_Custom Services

Introduction

Terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL) assay has been designed to detect apoptotic cells that undergo extensive DNA degradation during the late stages of apoptosis. This assay is based on the incorporation of modified dUTPs by the enzyme TdT at the 3’-hydroxyl termini of fragmented DNA, a hallmark as well as the ultimate determinate of apoptosis. The modifications are fluorophores or haptens, including biotin of bromine which can be detected directly in the case of a fluorescently-modified nucleotide, or indirectly with streptavidin or antibodies, if biotin-dUTP or BrdUTP are used.

TUNEL TUNEL

Procedures:

1. Deparaffinize sections in 2 changes of xylene, and then hydrate with 2 changes of ethanol.
2. Pretreatment: Use the proteinase K digestion method.
3. Peroxidase Blocking.
4. Pre-incubate the sections in TdT Reaction Buffer.
5. TdT Reaction.
6. Detection.
7. Counterstain with Gill’s hematoxylin.
8. Clear in xylene.

9. Coverslip with xylene based mounting medium.

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