苏州拓维生物技术有限公司

体外生物学

化合物洗脱实验

化合物洗脱实验可以帮助了解化合物与靶点的结合特性。在化合物洗脱实验中，首先用化合物处理细胞一定的时间，然后小心将化合物洗去，并换上新鲜培养基继续培养。最后选择合适的read-out来观察细胞的变化，如细胞增殖实验或者信号通路的变化等。通过比较不同化合物的洗脱实验结果，可以比较化合物与细胞内靶点结合的动力学，也有助于理解化合物在细胞水平的PK/PD关系。

引自：Edgar R. Wood et al. Cancer Res 2004;64:6652-6659

Recovery of EGFR autophosphorylation after treatment with GW572016, ZD-1839, and OSI-774. Logarithmically growing HN5 cells were treated with 1 μmol/L inhibitor in culture media for 4 hours. The media was removed, cells were washed twice, and fresh compound-free media was added. The cells were lysed at the indicated time after inhibitor washout, and EGFR was isolated by immunoprecipitation. A, tyrosine phosphorylated EGFR (pEGFR) was determined by Western blot using antiphosphotyrosine antibody. Lysates used for immunoprecipitation were prepared from cells treated with the following agent: Lane 1, DMSO control; Lane 2, GW572016; Lane 3, ZD-1839; and Lane 4, OSI-774. Total EGFR was measured from the same immunoprecipitate samples by Western blot using anti-EGFR antibody. B, The level of Tyrosine-phosphorylated EGFR was quantified for each condition and expressed as the percentage of vehicle-treated sample. The results represent the mean value of three independent experiments; bars, ±SD.

体外生物学

化合物洗脱实验

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体外生物学

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