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【文獻標題】Microcystin-LR influences the in vitro oocyte maturation of zebrafish by activating the MAPK pathway
【作者】Wanjing Liu, Chunhua Zhan, Tongzhou Zhang,et.al
【作者單位】華中農(nóng)業(yè)大學（Huazhong Agricultural University）
【文獻中引用產(chǎn)品】
斑馬魚卵黃原蛋白（VTG）ELISA試劑盒
【關鍵詞】Microcystin，Meiosis，Oocyte maturation，Hyper-phosphorylation，MAPK
【DOI】doi.org/10.1016/j.aquatox.2019.105261
【影響因子(IF)】7.2
【出版期刊】《Aquatic Toxicology》
【產(chǎn)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.

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