Titration-based normalization of antibody amount improves consistency of ChIP-seq experiments

Ariel Caride, Jin Sung Jang, Geng Xian Shi, Sam Lenz, Jian Zhong, Kwan Hyun Kim, Mariet Allen, Keith D. Robertson, Gianrico Farrugia, Tamas Ordog, Nilüfer Ertekin-Taner, Jeong Heon Lee

Research output: Contribution to journalArticlepeer-review


Chromatin immunoprecipitation (ChIP) is an antibody-based approach that is frequently utilized in chromatin biology and epigenetics. The challenge in experimental variability by unpredictable nature of usable input amounts from samples and undefined antibody titer in ChIP reaction still remains to be addressed. Here, we introduce a simple and quick method to quantify chromatin inputs and demonstrate its utility for normalizing antibody amounts to the optimal titer in individual ChIP reactions. For a proof of concept, we utilized ChIP-seq validated antibodies against the key enhancer mark, acetylation of histone H3 on lysine 27 (H3K27ac), in the experiments. The results indicate that the titration-based normalization of antibody amounts improves assay outcomes including the consistency among samples both within and across experiments for a broad range of input amounts.

Original languageEnglish (US)
Article number171
JournalBMC genomics
Issue number1
StatePublished - Dec 2023


  • Antibody titer
  • Chromatin immunoprecipitation
  • Chromatin quantification
  • Experimental consistency
  • H3K27ac

ASJC Scopus subject areas

  • Biotechnology
  • Genetics


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