Author:
Date:
2017-05-05
[Abstract] Preparing nuclei is necessary in a variety of experimental paradigms to study nuclear processes. In this protocol, we describe a method for rapid preparation of large number of relatively pure nuclei from Ascaris embryos or tissues that are ready to be used for further experiments such as chromatin isolation and ChIP-seq, nuclear RNA analyses, or preparation of nuclear extracts (Kang et al., 2016; Wang et al., 2016).
[摘要] 在各种实验范例中准备核是必要的,以研究核过程。在本协议中,我们描述了一种从准备用于进一步实验(如染色质分离和ChIP-seq,核RNA)的蛔虫胚胎或组织快速制备大量相对纯的核的方法分析或准备核提取物(Kang等人,2016; Wang等人,2016)。
背景 核分离通常是研究核事件的分子和生物化学方面的第一步。已经开发了几种方法来分离来自不同组织和细胞类型的细胞核。然而,除了C以外的线虫的细胞核分离方案很少。已经描述了线索,(Ooi等人,2010; Zanin等人,2011; Haenni等人,2012, )。已经使用寄生线虫蛔虫的胚胎来制备用于体外无细胞系统的各种提取物(Cohen等人,2004) ; Lall等人,2004)。然而,这些提取物通常是全细胞提取物。在这里,我们描述了从线虫蛔虫制备细胞核的方法。
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Author:
Date:
2017-04-05
[Abstract] Ovarian cancer is one of the most fatal tumors in women. Due to a lack of symptoms and adequate screening methods, patients are diagnosed at advanced stages with extensive tumor burden (Jelovac and Armstrong, 2011). Interestingly, ovarian cancer metastasis is generally found within the peritoneal cavity rather than other tissues (Lengyel, 2010; Tan et al., 2006). The reason behind this tissue tropism of the peritoneal cavity remains elusive. A prominent feature of this selectivity is ascites, the accumulation of fluid within the peritoneal cavity, containing, amongst others, immune cells, tumor cells and various soluble factors that can be involved in the progression of ovarian cancer (Kipps et al., 2013). The protocol described here is used to isolate mononuclear cells ...
[摘要] 卵巢癌是女性最致命的肿瘤之一。由于缺乏症状和适当的筛查方法,患者被诊断为具有广泛肿瘤负担的晚期阶段(Jelovac和Armstrong,2011)。有趣的是,卵巢癌转移通常发生在腹腔内,而不是其他组织(Lengyel,2010; Tan等人,2006)。腹膜腔组织向性背后的原因仍然难以捉摸。这种选择性的一个突出特征是腹水,腹膜内的液体积累,其中包含免疫细胞,肿瘤细胞和可参与卵巢癌进展的各种可溶性因子(Kipps等,。,2013)。这里描述的方案用于从腹水中分离单核细胞以研究腹膜腔内免疫系统的功能。
使用Lymphoprep 的梯度离心是分离外周血单核细胞(PBMC)的标准方案。我们稍微调整了方案,关于样品制备和洗涤步骤的数量,以便从腹水中分离单核细胞。
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