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Application of recombinant TAF3 PHD domain instead of anti-H3K4me3 antibody

机译：重组TAF3 PHD结构域代替抗H3K4me3抗体的应用

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Background Histone posttranslational modifications (PTMs) represent a focal point of chromatin regulation. The genome-wide and locus-specific distribution and the presence of distinct histone PTMs is most commonly examined with the application of histone PTM-specific antibodies. In spite of their central role in chromatin research, polyclonal antibodies suffer from disadvantages like batch-to-batch variability and insufficient documentation of their quality and specificity. Results To mitigate some of the pitfalls of using polyclonal antibodies against H3K4me3, we successfully validated the application of a recombinant TAF3 PHD domain as anti-H3K4me3 affinity reagent in peptide array, western blot and ChIP-like experiments coupled with qPCR and deep sequencing. Conclusions The successful addition of the TAF3 PHD domain to the growing catalog of recombinant affinity reagents for histone PTMs could help to improve the reproducibility, interpretation and cross-laboratory validation of chromatin data.
机译：背景组蛋白翻译后修饰（PTM）代表染色质调节的重点。应用组蛋白PTM特异性抗体最常检查全基因组范围和基因座特异性分布以及不同组蛋白PTM的存在。尽管多克隆抗体在染色质研究中起着核心作用，但仍存在批次间差异以及质量和特异性文献不足的缺点。结果为了缓解使用针对H3K4me3的多克隆抗体的一些陷阱，我们成功地验证了重组TAF3 PHD域作为抗H3K4me3亲和试剂在肽阵列，蛋白质印迹和ChIP样实验中与qPCR和深度测序相结合的应用。结论成功地将TAF3 PHD域添加到用于组蛋白PTM的重组亲和试剂中，可以帮助改善染色质数据的再现性，解释性和跨实验室验证。

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