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

ABSTRACT:

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.

SUBMITTER: Kungulovski G

PROVIDER: S-EPMC4802638 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature

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

Kungulovski Goran G Mauser Rebekka R Reinhardt Richard R Jeltsch Albert A

Epigenetics & chromatin 20160322

<h4>Background</h4>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.<h4>Results</h ...[more]

PMID: 27006701

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