Acetyl-Histone H3-K14 Rabbit Monoclonal Antibody (Q16695/P68431) for WB, ChIP, ELISA

Core Product Specifications and Parameters

Biological Background: Histone H3 Function and Localization

• Histone H3 is a core component of the nucleosome, the fundamental unit of chromatin, which packages DNA and regulates access for transcription, replication, repair, and chromosomal stability. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer (two each of H2A, H2B, H3, and H4).
• Acetylation at lysine 14 of Histone H3 (H3K14ac) is a well-characterized epigenetic mark associated with actively transcribed genes and open chromatin regions. It is catalyzed primarily by histone acetyltransferases such as GCN5/PCAF and is recognized by bromodomain-containing proteins. Related references: PMID:16457588, PMID:16185088, PMID:16267050, PMID:17194708
• Canonical Histone H3.1 (encoded by multiple genes including HIST1H3A, etc., SWISS P68431) is expressed in a replication-dependent manner during S phase and deposited genome-wide. In contrast, Histone H3.1t (SWISS Q16695, gene H3-4/HIST3H3) is a testis-specific variant that replaces canonical H3 during spermatogenesis.
• Both H3.1 and H3.1t share a high degree of sequence identity, including the N-terminal region containing K14. The antibody's immunogen, a synthetic peptide acetylated at K14, ensures specific recognition of this modification across both variants.
• Subcellular localization of Histone H3 is nuclear, where it is predominantly associated with chromosomes. Tissue specificity for H3.1t is restricted to testicular cells, while H3.1 is ubiquitous.

Experimental Guidance and Technical Tips

• Immunogen Design: The antibody was raised against a synthetic acetylated peptide around K14 of human H3, ensuring high specificity for the acetylated form. Competitive assays using non-acetylated peptide can be employed to confirm modification-specific binding.
• Western Blotting (WB): Histone H3 migrates at approximately 15 kDa under standard SDS-PAGE conditions. For optimal detection, consider using nuclear extracts and mild acid extraction to enrich histone proteins. Always include appropriate positive and negative controls (e.g., cells treated with histone deacetylase inhibitors vs. untreated).
• Chromatin Immunoprecipitation (ChIP): This antibody is listed for ChIP applications; however, optimization of crosslinking, chromatin shearing, and antibody concentration may be necessary depending on the target genomic regions and cell type.
• Immunofluorescence/Immunohistochemistry (IF/IHC): Nuclear staining is expected. For paraffin-embedded sections (IHC-P), antigen retrieval may be required. Validate antibody specificity with blocking peptides or by comparison with isotype controls.
• Species Reactivity: The antibody cross-reacts with human, mouse, and rat due to high sequence conservation around K14. For other species, verify sequence alignment before use.
• General: As a rabbit monoclonal antibody, it provides high affinity, batch-to-batch consistency, and low background. Store at -20°C and avoid repeated freeze-thaw cycles.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio equips diagnostic manufacturers, research laboratories, and academic institutes with a comprehensive selection of IVD raw materials, from validated antibody pairs and optimized monoclonal/polyclonal antibodies to bulk ancillary reagents. Our product portfolio includes specialized acetylated-antibody reagents like this Acetyl-Histone H3-K14 Rabbit mAb, which serves as a reliable tool for epigenetic biomarker development and chromatin research. Beyond standard catalog items, CamelBio offers custom antigen design and rare target raw-material sourcing to accelerate your assay development pipeline. Partner with CamelBio to streamline your product development from concept to clinic.