Proximity Ligation Assay to Quantify Foxp3 Acetylation in Regulatory T Cells.
Proximity Ligation Assay to Quantify Foxp3 Acetylation in Regulatory T Cells.
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DOI：
10.1007/978-1-4939-6527-4_21
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Beier UH
中科院分区：
文献类型：
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作者：
Jiao J;Han R;Hancock WW;Beier UH
通过免疫沉淀和免疫印迹确定蛋白质乙酰化可能具有挑战性，特别是如果感兴趣的组织数量较少，并且当无法获得高质量的乙酰化位点特异性抗体时。邻位连接试验允许一种灵敏的定量方法来评估调节性T细胞中Foxp3的乙酰化，在两天的周转时间内只需1.5 × 105个细胞。该方法在其他类似情况下具有潜在用途，当仅需要用少量样品并且在不存在可以评估感兴趣蛋白质中的翻译后修饰的特异性抗体的情况下确定感兴趣蛋白质的翻译后修饰时。
Determining protein acetylation by immunoprecipitation and immunoblotting can be challenging, especially if the tissue of interest is low in quantity, and when good quality acetylation-site specific antibodies are not available. Proximity ligation assays allow a sensitive and quantitative method to assess Foxp3 acetylation in regulatory T cells, with as little as 1.5 × 105 cells within two days turnaround time. This method is of potential use in other similar scenarios, when post-translational modifications of a protein of interest needs to be determined with only a small amount of sample and in the absence of specific antibodies that can assess the post-translational modification in the protein of interest.