TriMethyl-Histone H3-K4 (Q16695/P68431) Rabbit Monoclonal Antibody for ChIP-Seq, CUT&Tag, WB

Core Product Specifications and Parameters

Biological Background: Histone H3 Function and Localization

Histones are highly conserved basic proteins that serve as the core components of nucleosomes, the fundamental units of chromatin. Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Along with H2A, H2B, and H4, it forms the histone octamer around which approximately 147 base pairs of DNA are wrapped, compacting the genome and regulating accessibility to the transcriptional machinery.

The nucleosome dynamics, including post-translational modifications (PTMs) on histone tails, play a crucial role in epigenetic regulation. Histone H3 is subject to numerous modifications such as methylation, acetylation, phosphorylation, and more, which collectively constitute the "histone code" that governs chromatin structure and gene expression.

Among these PTMs, trimethylation of lysine 4 on histone H3 (H3K4me3) is a hallmark of active transcription. It is predominantly found at the promoters of actively transcribed genes and is involved in the recruitment of chromatin remodeling complexes and transcription factors. The antibody described here specifically recognizes the trimethylated form of H3K4. This modification is mediated by methyltransferases such as MLL complexes and is associated with gene activation. Related references: PMID:16267050 PMID:16457588 PMID:17194708

Histone H3.1t (encoded by H3-4 gene, accession Q16695) is a testis-specific variant of histone H3.1, expressed in testicular cells. The canonical histone H3.1 (accession P68431) is expressed in most tissues and is encoded by multiple genes. Both variants share high sequence similarity and are core components of the nucleosome.

Subcellular localization is exclusively nuclear, associated with chromosomes.

Experimental Guidance and Technical Tips

• This antibody was raised against a synthetic peptide corresponding to trimethylated lysine 4 of human Histone H3. For applications such as ChIP-seq, CUT&Tag, or IF, consider including a negative control using non-trimethylated peptide or an isotype control to confirm specificity.
• The antibody is predicted to have wide cross-reactivity due to the high conservation of the H3 N-terminal tail across species. However, validation in the relevant experimental system is recommended.
• For Western blotting, a band around 15 kDa corresponding to histone H3 should be detected. Ensure proper extraction of histones, particularly when analyzing methylation states.
• In ChIP assays, optimal shearing of chromatin and antibody titration should be determined empirically.
• For quantitative applications like ELISA or DB, consider using a standard curve with known amounts of trimethylated peptide.
• The antibody can be used in multiplexed assays with other histone modification antibodies to study the histone code combinatorially.

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As a comprehensive IVD raw-material supplier, CamelBio provides diagnostic manufacturers and research institutes with reliable access to high-quality antibodies like this TriMethyl-Histone H3-K4 Rabbit mAb. We support a wide range of needs for epigenetic research and clinical diagnostic assay development, offering validated antibody pairs, optimized monoclonal and polyclonal antibodies, bulk ancillary reagents, and custom sourcing for rare targets. Partner with CamelBio to streamline your raw-material procurement from discovery to commercialization.