Apoptotic cell detection in zebrafish embryos (TUNEL assay)

JP Jimin Park
GJ Gun Hyuk Jang
YJ Yeon Wook Jung
HS Hyunseon Seo
HH Hyung-Seop Han
JL Joonho Lee
YS Youngmin Seo
HJ Hojeong Jeon
MO Myoung-Ryul Ok
PC Pil-Ryung Cha
HS Hyun-Kwang Seok
KL Kwan Hyi Lee
YK Yu-Chan Kim
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DNA fragmentation was detected using the In Situ Cell Death Detection Kit, TMR red (Cat. No. 12156792910 Roche, Sigma, St. Louis, United States) according to the manufacturer’s protocol42,43. Embryos at 72 hpf were treated with solution, and alloys were fixed overnight at 4 °C in a 4% paraformaldehyde solution. Fixed embryos were washed three times with PBS and then were incubated in a permeabilization solution for 1 hour at room temperature. After washing with PBS, embryos were treated with TUNEL solution mixtures for 60 minutes at room temperature in the dark and then washed with PBS 3 times before optical observation.

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