HIST1H2BC (Ab-120) Antibody

What is HIST1H2BC (Ab-120) Antibody and what is its target protein?

HIST1H2BC (Ab-120) Antibody (PACO60634) is a polyclonal antibody produced in rabbits that specifically targets Histone H2B variant 1C (Hist1H2BC). This antibody recognizes a peptide sequence around site of Lysine 120 derived from Human Histone H2B type 1-C/E/F/G/I. Hist1H2BC is a key protein involved in chromatin structure and gene regulation, playing a crucial role in packaging DNA into chromatin and thereby influencing various cellular processes. The antibody exhibits high specificity and sensitivity towards human samples and serves as a valuable tool for investigating chromatin structure, gene expression regulation, and epigenetic modifications .

How does HIST1H2BC differ from other histone variants, and why is this significant for research?

Hist1H2BC belongs to the histone H2B family but has specific structural and functional characteristics that distinguish it from other variants. This specificity is important because different histone variants can have distinct roles in chromatin organization and gene regulation. Understanding these differences allows researchers to investigate specific epigenetic mechanisms and their implications in normal cellular processes and disease states. The dysregulation of Hist1H2BC has been implicated in various diseases, including cancer and developmental disorders, making it a significant target for research . When designing experiments, researchers should consider that while some antibodies like HIST1H2BC (Ab-12) target different epitopes (Lys-12) and have broader species reactivity (Human, Mouse, Rat), the (Ab-120) antibody is human-specific and targets the Lys-120 region .

What validated applications can HIST1H2BC (Ab-120) Antibody be used for?

The HIST1H2BC (Ab-120) Antibody has been validated for multiple research applications, primarily:

• Enzyme-Linked Immunosorbent Assay (ELISA): Validated at dilutions of 1:2000-1:10000

• Immunohistochemistry (IHC): Validated at dilutions of 1:10-1:100

The antibody has shown particularly promising results in IHC applications on paraffin-embedded human breast cancer tissue using a Leica BondTM system, following specific protocols involving dewaxing, hydration, and antigen retrieval under high pressure in citrate buffer (pH 6.0) . This versatility makes it suitable for researchers investigating the expression and localization of Hist1H2BC in various tissue samples and experimental conditions.

What are the optimal conditions for using HIST1H2BC (Ab-120) Antibody in IHC applications?

For optimal results in IHC applications, researchers should follow these methodological guidelines:

• Dilution range: Use the antibody at 1:10-1:100 dilution, with optimization recommended for specific tissue types

• Antigen retrieval: Perform high-pressure antigen retrieval in citrate buffer (pH 6.0)

• Blocking: Block sections with 10% normal goat serum to reduce background staining

• Incubation: Incubate sections with the antibody at the optimized dilution

• Detection system: Use an appropriate secondary antibody detection system compatible with rabbit IgG

Based on validated protocols, this antibody has shown excellent results in paraffin-embedded human breast cancer tissue using the Leica BondTM system . Researchers should consider that optimization may be required for different tissue types or experimental conditions.

How should HIST1H2BC (Ab-120) Antibody be stored and handled to maintain optimal activity?

To maintain optimal antibody activity and performance, researchers should adhere to these storage and handling recommendations:

• Storage conditions: Store the antibody at the recommended temperature in its original buffer

  • The antibody is supplied in liquid form containing preservative (0.03% Proclin 300)

  • Storage buffer consists of 50% Glycerol, 0.01M PBS, pH 7.4

  • Store at the temperature specified by the manufacturer (typically -20°C)

• Avoid repeated freeze-thaw cycles, which can degrade antibody performance

• When using the antibody, keep it on ice and return to storage promptly

• Follow recommended dilution protocols using appropriate buffers

• Consider aliquoting the stock solution to minimize freeze-thaw cycles

Proper storage and handling will help ensure consistent experimental results and extend the usable life of the antibody .

What controls should be included when designing experiments with HIST1H2BC (Ab-120) Antibody?

A robust experimental design with appropriate controls is essential for generating reliable results with HIST1H2BC (Ab-120) Antibody:

• Positive controls:

  • Human breast cancer tissue (validated target tissue)

  • Cell lines with known Hist1H2BC expression (e.g., HeLa cells)

• Negative controls:

  • Omission of primary antibody (technical negative control)

  • Tissues or cells with confirmed low/no expression of Hist1H2BC

  • Isotype control (rabbit IgG at the same concentration)

• Peptide blocking controls:

  • Pre-incubation of the antibody with the immunizing peptide to confirm specificity

• Cross-reactivity controls:

  • If investigating multiple histone variants, include controls to verify specificity against the target Hist1H2BC versus other histone variants

Including these controls will help validate the specificity of the antibody binding and authenticate experimental findings .

How does HIST1H2BC (Ab-120) Antibody compare to HIST1H2BC (Ab-12) Antibody for different research applications?

A direct comparison between these two antibodies reveals important differences that can guide experimental design decisions:

The choice between these antibodies should be based on the specific research application, species of interest, and target epitope relevance. For chromatin immunoprecipitation (ChIP) studies, the Ab-12 antibody would be preferable, while for human-specific IHC, the Ab-120 antibody might be more appropriate .

What advanced research applications can benefit from using HIST1H2BC (Ab-120) Antibody?

Beyond standard applications, this antibody can be valuable for several advanced research areas:

• Epigenetic modifications and chromatin structure studies:

  • Investigating post-translational modifications around Lys-120 of Hist1H2BC

  • Studying chromatin remodeling in different cell states or disease conditions

• Cancer research:

  • Analyzing Hist1H2BC expression patterns in various cancer types

  • Investigating its potential as a biomarker or therapeutic target

  • Studying its role in treatment response and resistance mechanisms

• Developmental biology:

  • Examining Hist1H2BC's role in cellular differentiation and development

  • Studying epigenetic reprogramming during development

• Multi-omics integration:

  • Combining with proteomics, genomics, or transcriptomics data to gain comprehensive insights into gene regulation

  • Correlating Hist1H2BC modification with gene expression profiles

These advanced applications leverage the specificity of the antibody to address complex research questions about chromatin regulation and gene expression dynamics .

Can HIST1H2BC (Ab-120) Antibody be adapted for high-throughput screening applications?

Yes, HIST1H2BC (Ab-120) Antibody can be adapted for high-throughput screening applications, though this requires specific methodological considerations:

• Integration into automated workflows:

  • Recent developments in cell-free expression and automated screening platforms can incorporate antibodies like HIST1H2BC (Ab-120) for high-throughput applications

  • The workflow can combine cell-free DNA template generation, protein synthesis, and binding measurements in an automated process

• Adaptation for AlphaLISA assays:

  • AlphaLISA (Amplified Luminescent Proximity Homogeneous Linked Immunosorbent Assay) enables rapid protein-protein interaction characterization without protein purification

  • This method is particularly valuable for high-throughput screening of antibody binding characteristics

• Miniaturization considerations:

  • Using robotic and acoustic liquid handling allows for highly parallel and miniaturized workflows

  • This approach enables a single researcher to express and profile the antigen-specific binding of hundreds of antibodies in 24 hours

• Implementation requirements:

  • Optimization of assay conditions for the specific antibody

  • Validation of signal-to-noise ratios in the high-throughput format

  • Development of appropriate controls and standards for data normalization

Researchers implementing such approaches should be aware that while full-length antibodies may require further optimization for high-throughput applications, synthetically dimerized antigen-binding fragments (sdFab) formats often show more consistent results in cell-free protein synthesis systems .

What are common issues encountered when using HIST1H2BC (Ab-120) Antibody in IHC and how can they be resolved?

Researchers may encounter several challenges when using this antibody in IHC applications:

• High background staining:

  • Problem: Non-specific binding resulting in excessive background

  • Solution: Increase blocking time/concentration (use 10% normal goat serum), optimize antibody dilution (try higher dilutions within the 1:10-1:100 range), reduce incubation time, or include additional washing steps

• Weak or absent staining:

  • Problem: Insufficient antigen detection

  • Solution: Optimize antigen retrieval conditions (ensure high-pressure retrieval in citrate buffer at pH 6.0), decrease antibody dilution (use lower dilutions within the recommended range), increase incubation time, or verify sample preparation and fixation protocols

• Variable staining intensity:

  • Problem: Inconsistent results between experiments

  • Solution: Standardize all protocol parameters (fixation time, antigen retrieval conditions, incubation times), prepare fresh working antibody dilutions for each experiment, and include positive controls in each batch

• Cross-reactivity:

  • Problem: Antibody binding to unintended targets

  • Solution: Perform peptide blocking controls, increase antibody dilution, optimize blocking conditions, or consider using more specific detection methods

• Edge artifacts:

  • Problem: Staining artifacts at tissue edges

  • Solution: Ensure proper tissue hydration, optimize antigen retrieval, use hydrophobic barriers around sections, and maintain consistent incubation conditions

Systematic optimization and the inclusion of appropriate controls are essential for troubleshooting these common issues .

How can researchers validate the specificity of HIST1H2BC (Ab-120) Antibody in their experimental system?

Validating antibody specificity is critical for generating reliable research data. Researchers should implement the following approaches:

• Western blot validation:

  • Run parallel Western blots with the antibody on relevant cell or tissue lysates

  • Confirm a single band at the expected molecular weight (~14 kDa for Hist1H2BC)

  • Compare with known positive and negative controls

• Peptide competition assay:

  • Pre-incubate the antibody with excess immunizing peptide before application

  • Observe elimination or significant reduction of signal in the presence of the peptide

• Genetic approaches:

  • Test the antibody on samples with genetic knockdown/knockout of Hist1H2BC

  • Observe reduced or absent signal in these samples

• Correlation with orthogonal methods:

  • Compare antibody staining patterns with RNA expression data (RT-qPCR or RNA-seq)

  • Use alternative antibodies targeting different epitopes of the same protein

  • Compare results with mass spectrometry-based protein identification

• Cross-reactivity assessment:

  • Test the antibody on tissues/cells expressing similar histone variants

  • Confirm the expected pattern of reactivity across species (primarily human for this antibody)

These validation steps will provide confidence in the specificity of the antibody and the reliability of experimental results .

What considerations are important when interpreting data generated using HIST1H2BC (Ab-120) Antibody?

Accurate data interpretation requires careful consideration of several factors:

• Expression pattern analysis:

  • Nuclear localization should be observed (consistent with histone proteins)

  • Compare staining patterns with known expression profiles of Hist1H2BC

  • Consider cell cycle-dependent expression patterns of histone proteins

• Technical limitations:

  • Recognize that the antibody primarily targets human samples, limiting cross-species applications

  • Acknowledge that polyclonal antibodies may show batch-to-batch variation

  • Consider potential post-translational modifications that might affect antibody binding

• Quantification approaches:

  • Develop standardized scoring systems for staining intensity and distribution

  • Use digital image analysis when possible for objective quantification

  • Include appropriate statistical analysis for comparative studies

• Biological context:

  • Interpret results in the context of known chromatin regulation mechanisms

  • Consider the influence of cell type, tissue context, and disease state on Hist1H2BC expression

  • Recognize that histone expression and modification are dynamic processes

• Integration with existing knowledge:

  • Compare findings with published literature on Hist1H2BC

  • Consider whether results align with known functions of H2B histones

  • Evaluate consistency with broader epigenetic regulation principles

How might HIST1H2BC (Ab-120) Antibody be utilized in emerging research areas such as single-cell analysis?

The antibody has potential applications in several cutting-edge research areas:

• Single-cell epigenetic profiling:

  • Adaptation for mass cytometry (CyTOF) to analyze Hist1H2BC expression at the single-cell level

  • Integration with single-cell Western blot technologies for protein expression analysis

  • Development of protocols for imaging mass cytometry to correlate Hist1H2BC distribution with spatial cellular context

• Multi-parameter imaging:

  • Combination with other epigenetic markers for multiplexed immunofluorescence

  • Integration with spatial transcriptomics to correlate histone modifications with gene expression patterns

  • Development of clearing techniques compatible with histone antibodies for 3D tissue imaging

• Liquid biopsy applications:

  • Detection of circulating nucleosomes containing Hist1H2BC in patient samples

  • Analysis of Hist1H2BC modifications in cell-free DNA/chromatin from biological fluids

  • Development of diagnostic or prognostic assays based on histone variant detection

• Rapid screening approaches:

  • Adaptation for miniaturized, high-throughput antibody screening platforms

  • Integration with AlphaLISA and other proximity-based detection methods

  • Incorporation into automated workflows for antibody validation and characterization

These emerging applications represent promising directions for expanding the utility of HIST1H2BC (Ab-120) Antibody in advanced research contexts .

What role might HIST1H2BC (Ab-120) Antibody play in investigating the connection between histone variants and disease states?

This antibody can serve as a valuable tool for exploring the relationship between Hist1H2BC and various pathological conditions:

• Cancer biology:

  • Investigation of Hist1H2BC expression changes across cancer types and stages

  • Analysis of correlations between Hist1H2BC modifications and treatment resistance

  • Exploration of its potential as a diagnostic or prognostic biomarker

  • Study of its role in chromatin organization changes during oncogenesis

• Neurodegenerative disorders:

  • Examination of histone variant distribution in neural tissues in disease states

  • Investigation of epigenetic dysregulation involving Hist1H2BC in neurodegeneration

  • Analysis of chromatin structure changes in affected brain regions

• Developmental disorders:

  • Study of Hist1H2BC's role in proper cellular differentiation and development

  • Investigation of epigenetic abnormalities in congenital conditions

  • Analysis of histone variant patterns in developmental timing regulation

• Inflammatory and immune disorders:

  • Examination of Hist1H2BC modifications in immune cell activation and differentiation

  • Study of chromatin changes during inflammatory responses

  • Investigation of histone variants as targets of autoantibodies in autoimmune conditions

The specificity of the antibody for the Lys-120 region makes it particularly valuable for research into post-translational modifications that might influence disease processes .

How can researchers integrate HIST1H2BC (Ab-120) Antibody data with other -omics approaches for comprehensive epigenetic analysis?

Integration of antibody-based data with multi-omics approaches can provide comprehensive insights into epigenetic regulation:

• Integration with genomics:

  • Combine ChIP-seq using HIST1H2BC (Ab-12) antibody (which is validated for ChIP) with RNA-seq to correlate Hist1H2BC localization with gene expression

  • Integrate with ATAC-seq to understand the relationship between histone variant incorporation and chromatin accessibility

  • Analyze together with DNA methylation data to explore the interplay between histone variants and DNA modifications

• Proteomics integration:

  • Correlate immunohistochemistry findings with mass spectrometry-based proteomics

  • Identify protein interaction networks involving Hist1H2BC using IP-MS approaches

  • Analyze post-translational modifications of Hist1H2BC using targeted proteomics

• Metabolomics connections:

  • Investigate relationships between metabolic states and histone modifications

  • Examine how metabolic changes affect Hist1H2BC incorporation and modification

• Computational integration:

  • Develop bioinformatic pipelines to integrate antibody-based imaging data with sequencing-based approaches

  • Apply machine learning algorithms to identify patterns across multi-omics datasets involving Hist1H2BC

  • Create predictive models for histone variant function based on integrated data

• Single-cell multi-omics:

  • Combine antibody-based detection with single-cell genomics/transcriptomics

  • Develop protocols for simultaneous analysis of histone variants and gene expression in individual cells

This integrated approach can provide a systems-level understanding of Hist1H2BC function in normal biology and disease contexts .