Simultaneous TUNEL and EdU Staining

EdU at a concentration of 3 μg/mL was added to the tissue slice culture medium 2 h before fixation. Simultaneous TUNEL and EdU staining was performed as described previously [35 (link)]. Briefly, tissue sections were deparaffinized in xylene followed by rehydration in graded alcohols and then blocked with PBS 3% bovine serum albumin (BSA). TUNEL reaction was performed using an In Situ Cell Death Detection Kit (Roche Life Sciences, Penzberg, Germany), after which the sections were incubated with Click-iT Alexa Fluor 594 (Invitrogen, Carlsbad, CA, USA) cocktail buffer for 30 min.

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Zhang W., Liao C.Y., Chtatou H., Incrocci L., van Gent D.C., van Weerden W.M, & Nonnekens J. (2019). Apalutamide Sensitizes Prostate Cancer to Ionizing Radiation via Inhibition of Non-Homologous End-Joining DNA Repair. Cancers, 11(10), 1593.

Publication 2019

In Situ Cell Death Detection Kit Click-iT Alexa Fluor 594

Alcohols Alexa 594 Bovine serum albumin Buffer Cell death Culture medium Rehydration Tissue Tunel Xylene

Corresponding Organization : Oncode Institute

Other organizations : Erasmus MC Cancer Institute

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Variable analysis

independent variables

EdU concentration (3 μg/mL)

dependent variables

Simultaneous TUNEL and EdU staining

control variables

Tissue slice culture medium
Incubation time (2 h before fixation)
Deparaffinization and rehydration procedure
Blocking with PBS 3% bovine serum albumin (BSA)
TUNEL reaction using In Situ Cell Death Detection Kit
Incubation with Click-iT Alexa Fluor 594 cocktail buffer (30 min)

controls

Positive control: Not specified
Negative control: Not specified

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