通过高效液相色谱法分离与Ras结合的[（32）P]标记核苷酸来定量绝对Ras-GDP/GTP水平。
Quantification of absolute Ras-GDP/GTP levels by HPLC separation of Ras-bound [(32)P]-labelled nucleotides.
作者信息
Rubio Ignacio, Pusch Rico, Wetzker Reinhard
机构信息
Research Unit Molecular Cell Biology, Medical Faculty, Friedrich-Schiller-University, Drackendorfer Str. 1, 07747 Jena, Germany.
出版信息
J Biochem Biophys Methods. 2004 Feb 27;58(2):111-7. doi: 10.1016/j.jbbm.2003.07.001.
Ras guanine nucleotide binding protein (GTPase) activation is a widely assessed readout in cell biological studies. We describe an improved approach for the quantitative analysis of total GDP and GTP bound to Ras. The present method involves HPLC separation and online detection/quantitation of Ras-bound [(32)P]-labelled GDP/GTP. As compared to standard approaches that are time consuming and/or provide only semi-quantitative data, this technique allows the rapid processing of large numbers of samples for the quantitative determination of Ras-bound GDP and GTP.
Ras鸟嘌呤核苷酸结合蛋白（GTP酶）激活是细胞生物学研究中广泛评估的一项指标。我们描述了一种用于定量分析与Ras结合的总GDP和GTP的改进方法。目前的方法包括HPLC分离以及对与Ras结合的[（32）P]标记的GDP/GTP进行在线检测/定量。与耗时且/或仅提供半定量数据的标准方法相比，该技术能够快速处理大量样品，用于定量测定与Ras结合的GDP和GTP。
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