检测 MYC 上的翻译后修饰。
Detection of Post-translational Modifications on MYC.
机构信息
Department of Molecular and Medical Genetics, Oregon Health and Science University, Portland, OR, USA.
Knight Cancer Institute, Oregon Health and Science University, Portland, OR, USA.
出版信息
Methods Mol Biol. 2021;2318:69-85. doi: 10.1007/978-1-0716-1476-1_5.
Detection of post-translational modifications in c-Myc is an invaluable tool in assessing Myc status, particularly in cancer. However, it can be challenging to detect these modifications. The evaluation of phosphorylation status of c-Myc can also be challenging with the current commercially available phosphorylation sensitive antibodies. Here, we describe protocols for the immunoprecipitation of endogenous c-Myc to probe for phosphorylation status, as well as the detection of ubiquitination and SUMOylation on c-Myc. We will also discuss the challenges of detecting phosphorylated c-Myc in formalin-fixed paraffin-embedded tissues by immunofluorescence and describe a protocol using a new rat monoclonal antibody we have generated suitable for this purpose.
检测 c-Myc 的翻译后修饰是评估 Myc 状态的宝贵工具，尤其是在癌症中。然而，检测这些修饰可能具有挑战性。目前市售的磷酸化敏感抗体也可能难以评估 c-Myc 的磷酸化状态。在这里，我们描述了用于免疫沉淀内源性 c-Myc 以探测磷酸化状态的方案，以及检测 c-Myc 上的泛素化和 SUMO 化的方案。我们还将讨论通过免疫荧光检测福尔马林固定石蜡包埋组织中磷酸化 c-Myc 的挑战，并描述我们为此目的生成的一种新的大鼠单克隆抗体的使用方案。
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