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DMEM, high glucose, GlutaMAXTM

公司名称: Thermo Fisher Scientific
产品编号: 61965026
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GFP-Grb2 Translocation Assay Using High-content Imaging to Screen for Modulators of EGFR-signaling
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2017-09-05
[Abstract]  High-content screening is a useful tool to understand complex cellular processes and to identify genes, proteins or small molecule compounds that modulate such pathways. High-content assays monitor the function of a protein or pathway by visualizing a change in an image-based readout, such as a change in the localization of a reporter protein. Examples of this can be the translocation of a fluorescently tagged protein from the cytoplasm to the nucleus or to the plasma membrane. One protein that is known to undergo such translocation is the Growth Factor Receptor-bound protein 2 (GRB2) that is ... [摘要]  高含量筛选是了解复杂细胞过程和鉴定调节这种途径的基因,蛋白质或小分子化合物的有用工具。高含量测定法通过显现基于图像的读数的变化来监测蛋白质或途径的功能,例如报告蛋白的定位的变化。其实例可以是将荧光标记的蛋白质从细胞质转移到细胞核或质膜。已知发生这种易位的一种蛋白质是生长因子受体结合蛋白2(GRB2),其在刺激生长因子受体并且随后经历内化后被招募到质膜。我们以前用GFP标记的Grb2来鉴定涉及EGFR信号的基因(Petschnigg等,2017)。最终,该测定可以适应于cDNA表达克隆(Freeman等人,2012),并且可用于早期药物发现以鉴定调节或抑制EGFR信号传导和内化的化合物(Antczak和Djaballah,2016)。
【背景】生长因子受体的信号转导对细胞维持正常功能至关重要,因此需要严格控制。生长因子受体的信号转导通过外部配体(例如,表皮生长因子,EGF)与跨膜受体(例如表皮生长因子受体(EGFR))和下游信号级联的活化(Yao等人,2015 )。 EGFR-信号传导的关键调节因子是生长因子受体结合蛋白2(Grb2),其由两个SH3结构域的内部SH2(Src同源性2)结构域组成。 Grb2通过其SH2结构域结合磷酸化酪氨酸残基上的活化生长因子受体,从而将受体活化与SOS-Ras-MAPK(丝裂原活化蛋白激酶)信号级联偶联。 ...

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