Soft Agar Anchorage-independent Assay
软琼脂锚着非依赖性分析
发布: 2013年10月20日第3卷第20期 DOI: 10.21769/BioProtoc.947 浏览次数: 14909
评审: Lin FangPinchas Tsukerman Fanglian He
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参见作者原研究论文
The authors used this protocol in:
Jan 2013
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Abstract
Chronic inflammation drives initiation of hepatocellular carcinoma (HCC), but the underlying mechanisms linking inflammation and tumor formation remain obscure. In this study, soft agar anchorage-independent assay were used to determine tumor transform activity of hepatoma cells with ISX over expression or knockdown in vitro.
Materials and Reagents
- HCC cells (Hep G2: ATCC® HB-8065TM and Hep 3B: ATCC® HB-8064TM)
- ISX fusion GFP expression plasmid or ISX shRNAi
- Agarose-LE (MDBio)
- 2x MEM no phenol red (Gibco)
- 100x NEAA (Gibco)
- FBS (Gibco)
- 10x PBS (MDBio)
- Crystal violet (Sigma-Aldrich, catalog number: C3866 )
- Methanol
- Ethanol
- ddH2O
Equipment
- 6 well culture dishes (Greiner Bio-One GmbH)
- Water bath
- Cell counter
- 37 °C incubator
Procedure
文章信息
版权信息
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Wang, L. and Hsu, S. (2013). Soft Agar Anchorage-independent Assay. Bio-protocol 3(20): e947. DOI: 10.21769/BioProtoc.947.
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分类
癌症生物学 > 通用技术 > 细胞生物学试验 > 细胞活性
细胞生物学 > 细胞染色 > 全细胞
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