【文獻標題】Fluoxastrobin-induced effects on acute toxicity, development toxicity,oxidative stress, and DNA damage in Danio rerio embryos

【作者】Cheng Zhang，Jingwen Zhang，Lusheng Zhu，et.al

【作者單位】山東農(nóng)業(yè)大學（Shandong Agricultural University）

【文獻中引用產(chǎn)品】

斑馬魚8羥基脫氧鳥苷（8-OHdG）ELISA試劑盒

【關鍵詞】96 h LC50，Delayed hatching，Teratogenicity，8-OHdG，AO-EB staining，Apoptosis

【DOI】doi.org/10.1016/j.scitotenv.2020.137069

【影響因子(IF)】5.92

【出版期刊】《Science of the Total Environment》

【產(chǎn)品原文引用】

2.7. Measurement of DNA damage induced by FLUO

To evaluate DNA damage to Danio rerio larvae induced by FLUO, 8-OHdG contents and apoptosis were monitored. The supernatant was prepared from the homogenates prepared in Section 2.5 after centrifugation at 12,857g and 4 °C for 10 min. The esultant solution was kept on ice during the entire procedure. A prior study (Guo et al., 2014)noted the significance of 8-OHdG in evaluating DNA damage. The 8-OHdH contents were monitored using an 8-OHdG ELISA kit (Hengyuan Biological Technology Co., Ltd., Shanghai, China) at 450 nm for 10 min (Multiskan MK3, Thermo Fisher Scientific, Massachusetts, USA) and in units of ng/g protein. The total protein contents were assessed as in Section 2.6.As per Sun et al. (2019) and Wang et al. (2018), apoptosis was monitored using acridine orange (AO)-ethidium bromide (EB) double dyeing kit (Solarbio Science & Technology Co., Beijing, China). At 96 hpf, ten larvae from each trial in Section 2.5 were rinsed twice with PBS (pH =7.4) and dyed with 20 μL of AO and EB working fluid (1/1, v/v) for 5 min and then rinsed twice with PBS. Tritricaine (MS-222) was used for 3 min to anesthetize larvae for better observation. Then, the apoptosis of larvae was observed and photographed under an inverted fluorescence microscope (xioVert. A1, Carl Zeiss AG, Oberkochen, Germany).

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