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

作者：Kungulovski Goran; Mauser Rebekka; Reinhardt Richard; Jeltsch Albert^*

来源：Epigenetics and Chromatin, 2016, 9(1): 11.

DOI：10.1186/s13072-016-0061-9

• Histone H3K4 methylation
• Reading domains
• Epigenetics
• Chromatin
• Antibody

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.

• 出版日期2016-3-22