【文献标题】Microcystin-LR influences the in vitro oocyte maturation of zebrafish by activating the MAPK pathway 【作者】Wanjing Liu1, Chunhua Zhan1, Tongzhou Zhang,et.al 【作者单位】华中农业大学（Huazhong Agricultural University） 【文献中引用产品】 斑马鱼卵黄原蛋白（VTG）ELISA试剂盒 【关键词】Microcystin，Meiosis，Oocyte maturation，Hyper-phosphorylation，MAPK 【DOI】https://doi.org/10.1016/j.aquatox.2019.105261 【影响因子(IF)】7.2 【出版期刊】《Aquatic Toxicology》 【产品原文引用】 PP2A ａnd MPF enzymatic activity ａnd VTG content PP2A enzymatic activity was assayed according to the instruction of Serine/Threonine Phosphatase Assay System (Promega, V2460;Madison, Wisconsin, USA). Live oocytes were rinsed ａnd homogenized in ice-cold PSB to obtain homogenate. The homogenate was added into the provided spin columns with Sephadex beads to remove endogenous phosphates. The supernatant was taken for detecting total soluble protein concentration using a bicinchoninic acid assay (BCA kit;Beyotime, Shanghai,China). The reaction was initiated by adding 15 μL of reaction premixes containing 5 μL of 1 mM phosphopeptide ａnd 10 μL of PPase-2A 5×reaction buffer. After reaction at 37 °C for 30 min,50 μL of stop solution was added to each well. The data was read at 630 nm. The activity levels of MPF ａnd the contents of VTG were determined through the zebrafish MPF ELISA Kit (Mlbio, Shanghai, China) ａnd the zebrafish VTG ELISA Kit (Huding, Shanghai, China), following the manufacturer’s protocol.
