HIST1H2BC (Ab-16) Antibody

Biological Significance of HIST1H2BC Protein

HIST1H2BC functions as a core component of nucleosomes, the fundamental units of chromatin structure that package and compact DNA. Nucleosomes critically regulate DNA accessibility to cellular machinery involved in transcription, replication, and repair processes. As part of the histone H2B family, HIST1H2BC plays a central role in maintaining proper chromatin organization and function .

The biological importance of HIST1H2BC extends beyond its structural role in chromatin organization. Recent research has uncovered that histone H2B degradation regulates insulin/IGF signaling pathways . Studies have shown that specific H2B isoforms exhibit differential stability during cellular stress conditions such as starvation, with most H2B isoforms becoming substantially reduced while certain variants maintain their levels . This selective degradation pattern suggests specialized roles for different H2B variants in stress response mechanisms.

Molecular Characteristics and Protein Structure

HIST1H2BC belongs to the highly conserved histone H2B family and is encoded by the HIST1H2BC gene. In databases such as UniProt, it is identified by accession number P62807 . The protein is also known by multiple synonyms including Histone H2B type 1-C/E/F/G/I, Histone H2B.1 A, H2B/a, Histone H2B.g, and several others, reflecting the complex nomenclature surrounding histone variants .

As a core histone protein, HIST1H2BC contains approximately 126 amino acids with a molecular weight of approximately 14 kDa, which is confirmed by Western blot analysis showing the observed band size matching the predicted size . The protein contains multiple sites for post-translational modifications, including the key lysine residue at position 16 that serves as the target epitope for the Ab-16 antibody .

Technical Specifications of HIST1H2BC (Ab-16) Antibody

The HIST1H2BC (Ab-16) Antibody (PACO60504) is a polyclonal antibody generated in rabbits using a synthetic peptide sequence surrounding the lysine residue at position 16 of the human Histone H2B type 1-C/E/F/G/I protein . This antibody preparation has been rigorously characterized to ensure specificity and reliability across multiple experimental applications.

Product Characteristics and Quality Control

The antibody is available as a 50μl liquid formulation preserved in 0.03% Proclin 300 with 50% glycerol in 0.01M PBS at pH 7.4 . This storage buffer provides stability and maintains antibody activity during shipping and storage. The antibody undergoes antigen affinity purification to ensure high specificity for the target epitope .

Quality control testing validates the antibody's performance across multiple applications:

• Western blotting (WB): Successfully detects HIST1H2BC in multiple human cell lines including HeLa, 293, A549, and HepG2, as well as in rat liver tissue, at a recommended dilution range of 1:100-1:1000 .

• Immunohistochemistry (IHC): Effectively stains human tissue sections including pancreatic tissue and cervical cancer specimens at recommended dilutions of 1:10-1:100 .

• Immunofluorescence (IF): Functions at dilutions of 1:1-1:10 for cellular localization studies .

• ELISA: Performs at higher dilutions of 1:2000-1:10000, demonstrating the antibody's high sensitivity .

Species Reactivity and Cross-Reactivity

The antibody shows confirmed reactivity with human and rat HIST1H2BC proteins . Western blot validation has verified specific binding to the target protein across multiple human cell lines and rat liver tissue . The antibody's species reactivity profile makes it suitable for comparative studies between human and rat model systems.

Research Applications and Methodological Considerations

The versatility of the HIST1H2BC (Ab-16) Antibody enables its application across diverse experimental approaches in chromatin biology and epigenetics research. Its utility spans from protein detection to localization studies and functional analyses.

Western Blot Applications

For Western blot applications, the antibody has been validated to detect a 14 kDa band corresponding to HIST1H2BC in various cell lysates . Optimal results are achieved at antibody concentrations of approximately 0.15μg/ml, with detection using a goat polyclonal anti-rabbit IgG secondary antibody at a 1:50000 dilution . This high dilution of secondary antibody indicates the primary antibody's strong binding affinity and specificity.

Western blot analysis has confirmed HIST1H2BC expression across multiple cell types, suggesting widespread expression of this histone variant in different tissues . This makes the antibody valuable for comparative studies of histone expression patterns across cell types and experimental conditions.

Immunohistochemistry and Immunofluorescence Applications

For IHC applications, the antibody has been successfully used on paraffin-embedded human tissue sections, including pancreatic tissue and cervical cancer specimens . The recommended protocol involves antigen retrieval under high pressure in citrate buffer (pH 6.0), followed by blocking with 10% normal goat serum and overnight incubation with the primary antibody at 4°C .

The ability to detect HIST1H2BC in both normal and cancer tissues makes this antibody valuable for investigating potential alterations in histone expression or localization in disease states. The validated IHC protocols provide researchers with a solid methodological foundation for tissue-based studies.

Comparison with Related Antibodies

The HIST1H2BC (Ab-16) Antibody targets the region surrounding lysine 16 of the HIST1H2BC protein. In contrast, the related HIST1H2BC (Ab-20) Antibody (PACO59665) targets the region surrounding lysine 20 . This difference in epitope targeting provides researchers with options for detecting different regions of the same protein, which can be valuable when studying post-translational modifications or protein interactions that might mask specific epitopes.

Distinctive Features of Ab-16 versus Ab-20

While both antibodies share several characteristics, including their polyclonal nature, rabbit host origin, and availability in 50μl liquid formulations, they differ in key aspects :

• Target epitope: Ab-16 targets lysine 16, while Ab-20 targets lysine 20 .

• Species reactivity: Ab-16 reacts with human and rat samples, whereas Ab-20 reacts with human and mouse samples .

• Validated applications: Ab-16 is validated for ELISA, WB, IHC, and IF, while Ab-20 is validated specifically for ELISA and WB .

These differences provide researchers with options when selecting the most appropriate antibody for their specific experimental system and research questions. The broader application range of Ab-16 makes it potentially more versatile for diverse experimental approaches.

Relevance to Disease Research and Epigenetics

Dysregulation of histone proteins, including HIST1H2BC, has been implicated in various pathological conditions, particularly cancer and developmental disorders . The ability to specifically detect and analyze HIST1H2BC using the Ab-16 antibody provides researchers with a valuable tool for investigating these disease associations.

Cancer Biology Applications

The validated use of the HIST1H2BC (Ab-16) Antibody in cancer tissues, such as cervical cancer specimens, demonstrates its utility for oncology research . Alterations in histone expression, modification, and localization are increasingly recognized as important factors in cancer development and progression. This antibody enables researchers to investigate potential roles of HIST1H2BC in these processes.

Recent research has shown that histone H2B variants can undergo selective degradation under stress conditions through polyubiquitination mechanisms . These degradation pathways may be dysregulated in cancer cells, affecting chromatin structure and gene expression patterns that contribute to malignant phenotypes.

Epigenetic Regulation Studies

Histones are central to epigenetic regulation through their post-translational modifications and interactions with chromatin-modifying enzymes. The HIST1H2BC (Ab-16) Antibody enables researchers to study the specific contribution of this histone variant to epigenetic mechanisms .

The antibody's ability to precisely target the region around lysine 16 is particularly relevant since histone lysine residues are frequent sites for modifications such as acetylation, methylation, and ubiquitination that constitute the "histone code" . These modifications regulate DNA accessibility and transcriptional activity, making their study essential for understanding gene regulation in normal development and disease states.