Acetyl-Histone H3 (Lys4) Rabbit Monoclonal Antibody for WB, ChIP, IF | Q16695/P68431

Core Product Specifications and Parameters

Biological Background: Histone H3 Function and Localization

• Histone H3 is a core component of the nucleosome, which wraps and compacts DNA into chromatin. This limits DNA accessibility to cellular machineries that require DNA as a template, thereby playing a central role in transcription regulation, DNA repair, DNA replication, and chromosomal stability. Related references: UniProtKB Q16695, UniProtKB P68431

• DNA accessibility is governed by a complex array of post-translational modifications (PTMs) on histones, collectively termed the "histone code," and nucleosome remodeling. These PTMs include acetylation, methylation, phosphorylation, citrullination, ADP-ribosylation, and various acylations. Related references: PMID:19608861, PMID:16267050, PMID:17194708

• Histone H3 localizes primarily to the nucleus and is associated with chromosomes throughout the cell cycle. Related references: UniProtKB Q16695, UniProtKB P68431

• While the canonical Histone H3.1 (encoded by multiple genes, e.g., HIST1H3A) is expressed broadly, the H3.1t variant (H3-4, HIST3H3) is expressed specifically in testicular cells, suggesting specialized roles in chromatin organization during spermatogenesis. Related references: UniProtKB Q16695

• Acetylation of lysine 4 on histone H3 (H3K4ac) is a well-characterized modification associated with active promoters and open chromatin. It often coexists with other activating marks and is read by bromodomain-containing proteins to facilitate transcriptional activation.

Experimental Guidance and Technical Tips

• The immunogen is a synthetic acetylated peptide corresponding to residues around K4 of human Histone H3. Due to the high conservation of the N-terminal tail among species, cross-reactivity with mouse and rat samples is predicted; however, it is advisable to verify performance in the specific experimental model.

• For ChIP assays, consider validating antibody specificity with acetylated and non-acetylated competitor peptides to confirm enrichment at target loci.

• In Western blotting, Histone H3 migrates at approximately 15 kDa. Optimize gel percentage and transfer conditions for low-molecular-weight proteins; nuclear or acid-extracted protein lysates are recommended for enriched signal.

• For immunofluorescence, test different fixation (e.g., 4% PFA vs. methanol) and permeabilization protocols, as these can influence epitope accessibility and background.

• Given the wide range of modifications at the H3 N-terminus, ensure that detection is not affected by neighboring PTMs by using appropriate controls and, if possible, mutant histones or recombinant proteins.

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