Histone H3.1 is a core component of nucleosomes, facilitating DNA compaction and transcriptional regulation. Antibodies against HIST1H3A are designed to detect specific post-translational modifications (PTMs) such as acetylation (e.g., K14ac) or methylation (e.g., K9me3), which influence gene expression and chromatin dynamics .
Specificity: Detects unmodified histone H3 (17 kDa) across human, mouse, and rat tissues .
Applications: Validated in WB (0.1–0.25 µg/ml), IHC (2–5 µg/ml), and Flow Cytometry (1–3 µg/1×10⁶ cells) .
Positive Controls:
Specificity: Recognizes endogenous histone H3 acetylated at K14 (19 kDa) .
Applications: Works in WB (1:500–1:2,000), IF (1:200–1:1,000), and ELISA .
Validation:
Some H3K14ac antibodies (e.g., Abcam ab82501) show unintended recognition of H3K36ac due to sequence similarity, as demonstrated by peptide microarray and competition assays .
Antibodies targeting H3K27me3 may cross-react with H3K4me3 in yeast (Saccharomyces cerevisiae), complicating bivalency studies .
Antibody binding can be influenced by adjacent modifications. For example, phosphorylation at Ser10 enhances 14-3-3 protein binding to H3K14ac in transcription activation assays .
IceChIP Validation: Anti-H3K27me3 antibodies (e.g., Abcam ab18521) show specificity in mouse embryonic stem cells, with signal loss in EED-null lines .
Spike-in Controls: Semi-synthetic nucleosomes validate antibody specificity under native and cross-linking conditions .
Cancer Research: Anti-H3K14ac antibodies detect acetylation changes in human colon adenocarcinoma and lung cancer tissues .