Glutaryl-HIST1H2BC (K116) Antibody

What is Glutaryl-HIST1H2BC (K116) and why is it significant in epigenetic research?

Glutaryl-HIST1H2BC (K116) refers to a specific post-translational modification (PTM) where a glutaryl group is attached to the lysine 116 residue of Histone H2B type 1-C/E/F/G/I protein. This modification is significant in epigenetic research because histone proteins play critical roles in packaging genomic DNA into nucleosomes and regulating gene expression . The nucleosome structure contains two subunits, each consisting of histones H2A, H2B, H3, and H4, with histone H1 serving as a junctional histone .

Glutarylation, like other histone PTMs (methylation, acetylation, phosphorylation, ubiquitination), serves as a signal for chromatin opening/compression and recruits factors that promote or antagonize transcription . Specifically, glutarylation at K116 of HIST1H2BC represents one of the newly discovered modifications that contributes to the "histone code," influencing gene regulation and potentially playing roles in various cellular processes and disease states.

The methodological approach to studying this modification typically begins with specific antibodies that recognize only the glutarylated form of this residue, allowing researchers to investigate its presence, distribution, and function in different cellular contexts .

How does Glutaryl-HIST1H2BC (K116) Antibody differ from antibodies targeting other histone modifications?

Glutaryl-HIST1H2BC (K116) Antibody is specifically designed to recognize and bind to histone H2B type 1-C/E/F/G/I that has been modified with a glutaryl group at the lysine 116 position . This specificity distinguishes it from antibodies targeting other histone modifications in several key ways:

• Epitope specificity: This antibody recognizes only the glutaryl modification at the K116 position, not other positions or modifications like acetylation or methylation .

• PTM chemistry: Glutarylation involves the addition of a five-carbon dicarboxylic acid derivative, which creates a structurally distinct modification compared to smaller modifications like acetylation (two-carbon) or larger ones like ubiquitination .

• Research applications: While many histone modification antibodies are validated for multiple applications, the Glutaryl-HIST1H2BC (K116) Antibody has been specifically validated for ELISA, Western blot, and immunofluorescence techniques .

Methodologically, when selecting between different modification-specific antibodies, researchers should consider the following approach:

• Verify the exact epitope recognition (position and modification type)

• Check cross-reactivity data with similar modifications

• Review validation data for your specific application

• Consider using complementary antibodies to different modifications in parallel experiments to build a comprehensive picture of the histone modification landscape .

What are the recommended experimental conditions for using Glutaryl-HIST1H2BC (K116) Antibody in Western blot applications?

For optimal Western blot results with Glutaryl-HIST1H2BC (K116) Antibody, the following methodological approach is recommended:

Sample Preparation:

• Extract histones using acid extraction methods (0.2N HCl or 0.4N H₂SO₄) to preserve post-translational modifications

• Include deacetylase and protease inhibitors in extraction buffers

• Load 10-20 μg of histone extracts or 50-100 μg of whole cell lysates per lane

Western Blot Protocol:

• Dilution range: Use antibody at 1:100-1:1000 dilution for Western blot applications

• Blocking: 5% non-fat dry milk or BSA in TBST for 1 hour at room temperature

• Primary antibody incubation: Overnight at 4°C in blocking buffer

• Washing: 3-5 times with TBST, 5 minutes each

• Secondary antibody: Anti-rabbit IgG (the antibody is raised in rabbits)

• Detection system: ECL or similar chemiluminescence system

Controls:

• Positive control: Human cell lines with known glutarylation (HeLa, HEK293)

• Negative control: Samples treated with glutarylation-specific eraser enzymes

• Loading control: Total H2B or other core histones

When troubleshooting, consider that the high specificity of this antibody means that signal intensity will directly correlate with the abundance of glutarylation at the specific K116 site, which may vary significantly across different cell types and physiological conditions .

How can Glutaryl-HIST1H2BC (K116) Antibody be used to investigate the relationship between metabolism and epigenetic regulation?

Glutarylation is metabolically linked to the TCA cycle and lysine metabolism, making Glutaryl-HIST1H2BC (K116) Antibody a valuable tool for investigating metabolism-epigenetics connections. A comprehensive methodological approach includes:

Experimental Design:

• Metabolic perturbation experiments:

  • Treat cells with glutaryl-CoA precursors or inhibitors

  • Modify cellular energy status (glucose deprivation, fatty acid supplementation)

  • Induce mitochondrial dysfunction using specific inhibitors

• Multi-omics integration:

  • Combine ChIP-seq using Glutaryl-HIST1H2BC (K116) Antibody with:

    • Metabolomics to measure TCA cycle intermediates

    • RNA-seq to assess transcriptional consequences

    • Proteomics to identify associated protein complexes

• Enzyme manipulation studies:

  • Overexpress or knockdown glutaryl-lysine transferases

  • Modulate deglutarylase enzymes (typically sirtuins)

  • Assess changes in glutarylation patterns using the antibody

Data Analysis Framework:

• Track changes in glutarylation levels under different metabolic conditions

• Correlate glutarylation with gene expression changes

• Map glutarylation sites to chromatin regions with specific functions

This methodological approach allows researchers to establish direct connections between cellular metabolism, histone glutarylation at K116, and transcriptional outcomes, potentially revealing new regulatory mechanisms in metabolic diseases .

What are the optimal conditions for Chromatin Immunoprecipitation (ChIP) using Glutaryl-HIST1H2BC (K116) Antibody?

While standard ChIP protocols provide a foundation, optimizing ChIP with Glutaryl-HIST1H2BC (K116) Antibody requires specific methodological considerations:

Pre-ChIP Considerations:

• Cell number: Start with 1-5×10⁶ cells for targeted ChIP; 10-20×10⁶ for ChIP-seq

• Crosslinking: 1% formaldehyde for 10 minutes at room temperature

• Include glutarylation preservatives: Add 5-10 mM sodium butyrate and KDAC inhibitors

ChIP Protocol Optimization:

• Chromatin shearing:

  • Target fragments of 200-500 bp for optimal resolution

  • Sonication parameters: 10-15 cycles (30s ON/30s OFF) at medium power

  • Verify fragmentation by agarose gel electrophoresis

• Immunoprecipitation:

  • Antibody amount: 2-5 μg per ChIP reaction

  • Pre-clearing: 1-2 hours with protein A/G beads

  • Incubation time: Overnight at 4°C with gentle rotation

  • Washing stringency: Gradually increase salt concentration (150-500 mM NaCl)

• Controls:

  • Input control: 5-10% of chromatin before immunoprecipitation

  • IgG control: Same amount of rabbit IgG as the primary antibody

  • Positive control: Antibody against total H2B or H3

  • Glutarylation-depleted control: Samples treated with deglutarylases

Analysis Recommendations:

• For ChIP-qPCR: Design primers for regions with known or suspected glutarylation involvement

• For ChIP-seq: Use specialized peak calling algorithms sensitive to histone modification patterns

• Compare glutarylation patterns with other histone marks to identify co-occurrence or mutual exclusion

This optimized approach enhances sensitivity and specificity when mapping glutarylation at K116 across the genome, providing insight into its regulatory functions .

How can researchers distinguish between glutarylation at K116 versus other lysine residues on HIST1H2BC?

Distinguishing between glutarylation at different lysine residues requires a methodological approach combining antibody-based detection with advanced analytical techniques:

Antibody Specificity Verification:

• Peptide competition assay:

  • Pre-incubate the Glutaryl-HIST1H2BC (K116) Antibody with:

    • Glutarylated K116 peptide (should eliminate signal)

    • Glutarylated non-K116 peptides (should not affect signal)

    • Unmodified K116 peptide (should not affect signal)

  • Proceed with Western blot or immunofluorescence

  • Loss of signal only with the glutarylated K116 peptide confirms specificity

• Cross-reactivity testing:

  • Compare signal patterns using:

    • Glutaryl-HIST1H2BC (K116) Antibody (PACO60520)

    • Glutaryl-HIST1H2BC (K120) Antibody (PACO60524)

    • Pan-glutaryl-lysine antibodies

Orthogonal Validation Techniques:

• Mass spectrometry approach:

  • Digest histones with trypsin or alternative proteases

  • Enrich for glutarylated peptides using anti-glutaryl-lysine antibodies

  • Perform LC-MS/MS analysis with high mass accuracy

  • Identify specific glutarylation sites based on mass shifts

  • Quantify relative abundance of modifications at different sites

• Site-directed mutagenesis:

  • Generate K116R, K120R, and other lysine-to-arginine mutants

  • Express in cells and immunoblot with site-specific antibodies

  • Signal loss with the specific mutation confirms antibody specificity

Data Integration Framework:

• Create a glutarylation site map across the HIST1H2BC protein

• Compare relative abundances of glutarylation at different sites

• Correlate site-specific glutarylation with functional outcomes

This comprehensive approach ensures accurate identification and functional characterization of glutarylation specifically at the K116 position versus other sites .

What are the most common challenges when working with Glutaryl-HIST1H2BC (K116) Antibody and how can they be addressed?

Researchers working with Glutaryl-HIST1H2BC (K116) Antibody commonly encounter several challenges that require specific methodological solutions:

Challenge 1: Low Signal Intensity

• Causes:

  • Low abundance of glutarylation at K116

  • Glutarylation loss during sample preparation

  • Suboptimal antibody concentration

• Methodological Solutions:

  • Treat cells with glutarylation enhancers (glutarate or glutaryl-CoA precursors)

  • Add deglutarylase inhibitors during all steps of sample preparation

  • Optimize antibody concentration (test range from 1:100 to 1:1000 for WB)

  • Increase protein loading (up to 50-100 μg for whole cell lysates)

  • Extend primary antibody incubation time (overnight at 4°C)

  • Use high-sensitivity detection systems (enhanced chemiluminescence)

Challenge 2: High Background or Non-specific Binding

• Causes:

  • Insufficient blocking

  • Cross-reactivity with other glutarylated proteins

  • Secondary antibody issues

• Methodological Solutions:

  • Optimize blocking (test 3-5% BSA vs. non-fat milk)

  • Increase washing duration and number of washes

  • Use highly purified antibody preparations

  • Perform peptide competition controls to verify specificity

  • Reduce secondary antibody concentration

  • Include 0.1-0.3% Triton X-100 in wash buffers

Challenge 3: Inconsistent Results Across Experiments

• Causes:

  • Variability in glutarylation levels under different conditions

  • Technical variations in sample preparation

  • Lot-to-lot antibody variations

• Methodological Solutions:

  • Standardize cell culture conditions (passage number, confluence)

  • Create standard operating procedures for histone extraction

  • Include internal controls in each experiment

  • Test and validate each new antibody lot

  • Generate standard curves using known glutarylated samples

Challenge 4: Compatibility with Other Techniques

• Causes:

  • Fixation effects on epitope accessibility

  • Buffer incompatibility

  • Antibody format limitations

• Methodological Solutions:

  • For IF: Test different fixation methods (4% PFA vs. methanol)

  • For ChIP: Optimize crosslinking conditions

  • For flow cytometry: Test permeabilization protocols

  • Consider using secondary antibody conjugates optimized for specific applications

This systematic troubleshooting approach helps researchers overcome technical challenges to obtain reliable and reproducible results when studying glutarylation at K116 of HIST1H2BC .

How can multiplexed detection of histone modifications be performed to study the interplay between glutarylation and other modifications?

Multiplexed detection of histone modifications provides insight into the complex interplay between glutarylation and other epigenetic marks. A comprehensive methodological approach includes:

Sequential Immunoblotting Strategy:

• Strip-and-reprobe technique:

  • Perform initial Western blot with Glutaryl-HIST1H2BC (K116) Antibody

  • Document results thoroughly

  • Strip membrane (mild stripping buffer: 200mM glycine, 0.1% SDS, 1% Tween-20, pH 2.2)

  • Verify complete stripping with secondary antibody only

  • Reprobe with antibodies against other modifications

  • Typical order: start with lowest abundance modification and proceed to higher abundance

• Parallel blotting approach:

  • Run identical samples on multiple gels

  • Transfer and probe each membrane with different modification-specific antibodies

  • Align images based on molecular weight markers and loading controls

Multiplex Immunofluorescence Methods:

• Sequential immunostaining:

  • Use primary antibodies from different host species

  • Apply Glutaryl-HIST1H2BC (K116) Antibody (rabbit)

  • Apply second modification antibody (mouse or goat)

  • Use species-specific secondary antibodies with distinct fluorophores

  • Include nuclear counterstain (DAPI)

• Advanced multiplexing techniques:

  • Tyramide signal amplification for sequential staining with same-species antibodies

  • Spectral unmixing for closely overlapping fluorophores

  • Antibody stripping and restaining protocols

Mass Spectrometry-Based Approach:

• Bottom-up histone analysis:

  • Digest histones with appropriate proteases

  • Analyze by LC-MS/MS with data-dependent acquisition

  • Identify peptides carrying multiple modifications

  • Quantify relative abundances of different modification combinations

• Middle-down approach:

  • Use GluC or AspN to generate larger histone fragments

  • Analyze by high-resolution MS

  • Map combinatorial modification patterns

Data Integration and Visualization:

• Generate modification co-occurrence matrices

• Apply machine learning algorithms to identify modification patterns

• Create visual maps of histone modification networks

This comprehensive multiplexing approach allows researchers to determine whether glutarylation at K116 co-occurs with, or is mutually exclusive of, other histone modifications, providing insight into the functional histone code .

What are the latest advanced applications of Glutaryl-HIST1H2BC (K116) Antibody in disease research?

The Glutaryl-HIST1H2BC (K116) Antibody is increasingly employed in cutting-edge disease research applications that leverage methodological innovations:

Cancer Research Applications:

• Patient-derived xenograft (PDX) models:

  • Use the antibody to track glutarylation changes during tumor progression

  • Methodological approach:

    • Establish PDX models from patient biopsies

    • Monitor glutarylation at different stages using immunohistochemistry

    • Correlate with disease progression and treatment response

    • Create predictive biomarker panels combining glutarylation with other markers

• Drug discovery pipelines:

  • Screen compounds affecting glutarylation enzymes

  • Methodological approach:

    • High-throughput screening with automated Western blots

    • Quantify glutarylation changes using image analysis software

    • Establish dose-response relationships

    • Validate hits in cellular and animal models

Neurodegenerative Disease Research:

• Brain region-specific epigenetic mapping:

  • Map glutarylation patterns across brain regions

  • Methodological approach:

    • Immunohistochemistry on brain sections

    • Compare glutarylation in affected vs. unaffected regions

    • Correlate with neuronal function and pathology

    • Integrate with other epigenetic marks in three-dimensional chromatin maps

• Single-cell epigenomics:

  • Analyze glutarylation heterogeneity in neuronal populations

  • Methodological approach:

    • Single-cell Western blot or CyTOF with the antibody

    • Identify cell type-specific glutarylation signatures

    • Correlate with disease susceptibility

Metabolic Disease Innovations:

• Circadian rhythm studies:

  • Track glutarylation changes throughout the day

  • Methodological approach:

    • Time-course sampling and Western blot analysis

    • ChIP-seq at different time points

    • Correlate with metabolic fluctuations and gene expression

• Nutrigenomic applications:

  • Investigate dietary influences on histone glutarylation

  • Methodological approach:

    • Dietary interventions followed by histone analysis

    • Compare different dietary components' effects on glutarylation

    • Link to metabolic outcomes and gene expression changes

Emerging Technologies:

• Spatial transcriptomics integration:

  • Combine glutarylation detection with spatial gene expression

  • Methodological approach:

    • Perform immunofluorescence with the antibody

    • Follow with in situ RNA hybridization

    • Create spatial maps correlating glutarylation with gene expression

• CRISPR epigenome editing:

  • Target glutarylation machinery to specific genomic loci

  • Methodological approach:

    • Design dCas9-glutaryltransferase fusions

    • Verify targeted glutarylation using ChIP with the antibody

    • Assess functional consequences of site-specific glutarylation

These advanced applications demonstrate how Glutaryl-HIST1H2BC (K116) Antibody enables researchers to explore the role of histone glutarylation in disease pathogenesis and potential therapeutic interventions .

What are the recommended protocols for using Glutaryl-HIST1H2BC (K116) Antibody in immunofluorescence applications?

For optimal immunofluorescence results with Glutaryl-HIST1H2BC (K116) Antibody, researchers should follow this detailed methodological protocol:

Sample Preparation:

• Cell culture preparation:

  • Grow cells on coverslips or chamber slides to 70-80% confluence

  • Consider synchronizing cells if studying cell-cycle dependent changes

• Fixation options:

  • Primary method: 4% paraformaldehyde for 15 minutes at room temperature

  • Alternative method: Ice-cold methanol for 10 minutes at -20°C

  • Note: Test both methods as epitope accessibility may vary

• Permeabilization:

  • Use 0.1-0.2% Triton X-100 in PBS for 10 minutes

  • For nuclear-specific staining, ensure complete permeabilization

Immunostaining Protocol:

• Blocking:

  • Block with 3-5% BSA or normal serum in PBS for 1 hour at room temperature

  • Include 0.1% Tween-20 to reduce background

• Primary antibody incubation:

  • Dilute Glutaryl-HIST1H2BC (K116) Antibody at 1:1-1:10 in blocking buffer

  • Incubate overnight at 4°C in a humidified chamber

  • For co-staining, apply compatible antibodies simultaneously or sequentially

• Washing:

  • Wash 3 times with PBS containing 0.05% Tween-20, 5 minutes each

  • Ensure thorough washing to minimize background

• Secondary antibody:

  • Apply anti-rabbit secondary antibody (fluorophore-conjugated)

  • Incubate for 1-2 hours at room temperature

  • Protect from light to prevent photobleaching

• Counterstaining:

  • Nuclear counterstain: DAPI (1 μg/ml) for 5 minutes

  • Optional: Add phalloidin for F-actin visualization

• Mounting:

  • Mount with anti-fade mounting medium

  • Seal edges with nail polish for long-term storage

Controls and Validation:

• Positive control: Human cell lines with known glutarylation

• Negative controls:

  • Primary antibody omission

  • Peptide competition (pre-incubate antibody with immunizing peptide)

• Specificity control: Compare with other glutarylation site antibodies

Image Acquisition and Analysis:

• Microscopy settings:

  • Capture images with confocal or wide-field fluorescence microscopy

  • Use consistent exposure settings across samples

  • Z-stack acquisition for 3D analysis of nuclear distribution

• Quantification methods:

  • Measure nuclear intensity using ImageJ or similar software

  • Perform co-localization analysis with other histone marks

  • Consider single-cell analysis to detect population heterogeneity

This comprehensive protocol ensures reliable detection and quantification of HIST1H2BC glutarylation at K116 in cellular contexts, allowing for spatial analysis of this epigenetic mark .

How can researchers quantitatively analyze Western blot data for Glutaryl-HIST1H2BC (K116) across different experimental conditions?

Quantitative analysis of Glutaryl-HIST1H2BC (K116) Western blot data requires a rigorous methodological approach to ensure accuracy and reproducibility:

Experimental Design for Quantification:

• Sample preparation controls:

  • Include biological replicates (minimum n=3)

  • Prepare all samples simultaneously with identical protocols

  • Include a concentration gradient of positive control samples

  • Ensure equal protein loading across all wells

• Loading and normalization strategy:

  • Always run total H2B on the same blot or on a parallel blot

  • Include loading controls (β-actin, GAPDH, or total protein stain)

  • Consider dual-channel Western blot systems for simultaneous detection

Image Acquisition Protocol:

• Optimal image capture:

  • Use a digital imaging system with linear dynamic range

  • Avoid oversaturation (check histogram during acquisition)

  • Capture multiple exposures if signal intensity varies greatly

  • Include molecular weight markers in all images

• File handling:

  • Save raw, unmodified files in original format

  • Use lossless compression formats for analysis (TIFF preferred)

  • Maintain consistent image resolution across experiments

Quantification Methods:

• Densitometry analysis workflow:

  • Define regions of interest (ROIs) of consistent size

  • Subtract local background for each lane

  • Measure integrated density or mean gray value

  • Normalize glutarylation signal to total H2B or loading control

  • Calculate relative changes compared to control conditions

• Advanced quantification approaches:

  • Time-course analysis: Plot glutarylation changes over time

  • Dose-response relationship: Plot glutarylation vs. treatment concentration

  • Signal ratio analysis: Compare glutarylation at different sites

Statistical Analysis Framework:

• Basic statistical tests:

  • Paired or unpaired t-tests for two-group comparisons

  • ANOVA with appropriate post-hoc tests for multiple groups

  • Non-parametric tests if normality assumptions are violated

• Data presentation guidelines:

  • Include representative blot images with molecular weight markers

  • Present quantification data as bar charts with error bars (SEM or SD)

  • Indicate statistical significance levels (* p<0.05, ** p<0.01, etc.)

  • Include sample size (n) in figure legends

Example Quantification Table Format:

This systematic approach to quantitative Western blot analysis ensures reliable measurement of Glutaryl-HIST1H2BC (K116) levels, enabling meaningful comparison across experimental conditions and accurate interpretation of biological significance .

How can researchers integrate epigenomic data from Glutaryl-HIST1H2BC (K116) ChIP-seq with transcriptomic and other multi-omics datasets?

Integrating epigenomic data from Glutaryl-HIST1H2BC (K116) ChIP-seq with other omics datasets requires a sophisticated methodological approach:

Data Generation and Quality Control:

• Multi-omics experimental design:

  • Use matched samples for all omics analyses

  • Include appropriate replicates (minimum triplicate)

  • Process all samples in parallel when possible

  • Incorporate spike-in controls for quantitative comparisons

• ChIP-seq specific considerations:

  • Generate input controls from the same chromatin

  • Include IgG controls to assess non-specific binding

  • Consider using spike-in normalization (e.g., Drosophila chromatin)

  • Sequence to sufficient depth (>20 million uniquely mapped reads)

Computational Analysis Pipeline:

• Primary data processing:

  • ChIP-seq: Alignment, peak calling, signal normalization

  • RNA-seq: Alignment, quantification, differential expression analysis

  • ATAC-seq/DNase-seq: Accessibility profiling

  • Proteomics: Protein identification and quantification

• Integration analysis methods:

  • Correlation analysis:

    • Calculate correlation between glutarylation signal and gene expression

    • Generate heatmaps of multi-omics data centered on glutarylation peaks

    • Perform k-means clustering to identify co-regulated gene sets

  • Feature overlapping:

    • Define genomic intervals (promoters, enhancers, gene bodies)

    • Calculate glutarylation enrichment in these regions

    • Compare with other histone marks and chromatin features

  • Advanced statistical integration:

    • Factor analysis for quantification of latent variables

    • Bayesian network modeling for causal relationship inference

    • Machine learning for pattern recognition and prediction

Visualization Strategies:

• Genome browser tracks:

  • Display aligned ChIP-seq data alongside RNA-seq, ATAC-seq

  • Create custom tracks for different experimental conditions

  • Generate aggregate plots around genomic features

• Integrated visualization tools:

  • Circos plots for genome-wide interactions

  • Network diagrams showing relationships between multiple data types

  • Principal component analysis plots for global data structure

Functional Interpretation Framework:

• Pathway enrichment analysis:

  • Identify biological processes enriched in glutarylated regions

  • Compare with pathways from differentially expressed genes

  • Integrate with metabolic pathway analysis if metabolomics data available

• Motif analysis:

  • Identify transcription factor binding motifs enriched near glutarylation sites

  • Correlate with transcription factor expression data

  • Test for co-occurrence with other histone modifications

• Systems biology approach:

  • Construct gene regulatory networks incorporating glutarylation data

  • Identify key nodes and regulatory hubs

  • Simulate perturbations to predict functional outcomes

Example Multi-omics Integration Workflow:

• Perform ChIP-seq with Glutaryl-HIST1H2BC (K116) Antibody

• Conduct RNA-seq on matched samples

• Optional: Include ATAC-seq, other histone mark ChIP-seq, metabolomics

• Process each dataset through appropriate pipelines

• Identify glutarylation-enriched regions and associated genes

• Correlate glutarylation patterns with gene expression changes

• Perform pathway analysis on correlated gene sets

• Validate key findings with targeted experiments

This comprehensive integration approach enables researchers to situate glutarylation within the broader epigenetic landscape and understand its functional consequences on gene expression and cellular phenotypes .

What are the emerging research areas and future directions for studies using Glutaryl-HIST1H2BC (K116) Antibody?

Glutaryl-HIST1H2BC (K116) Antibody is poised to play a critical role in several emerging research directions that promise to expand our understanding of epigenetic regulation:

Single-Cell Epigenomics:

• Methodological approach:

  • Adapt ChIP protocols for single-cell applications using Glutaryl-HIST1H2BC (K116) Antibody

  • Combine with single-cell RNA-seq for correlated epigenetic-transcriptomic profiles

  • Develop computational frameworks to analyze cell-to-cell variation in glutarylation

  • Potential to reveal heterogeneity in epigenetic regulation previously masked in bulk analyses

Dynamic Regulation of Glutarylation:

• Methodological approach:

  • Develop real-time imaging systems using fluorescent antibody derivatives

  • Create glutarylation biosensors for live-cell monitoring

  • Employ rapid induction systems to track glutarylation kinetics

  • Apply mathematical modeling to understand dynamic glutarylation regulation

Cross-talk with Other Acylations:

• Methodological approach:

  • Perform multiplexed detection of different acylations (acetylation, butyrylation, glutarylation)

  • Map modification co-occurrence and exclusivity patterns

  • Identify shared and specific regulatory enzymes

  • Develop targeted approaches to manipulate specific acylations independently

Therapeutic Targeting of Glutarylation Pathways:

• Methodological approach:

  • Screen for small molecules affecting glutarylation levels

  • Develop glutarylation-specific reader, writer and eraser modulators

  • Use Glutaryl-HIST1H2BC (K116) Antibody to validate target engagement

  • Apply in disease models to assess efficacy of epigenetic therapies

Evolutionary Conservation of Glutarylation:

• Methodological approach:

  • Test antibody cross-reactivity with model organisms

  • Compare glutarylation patterns across species

  • Identify conserved regulatory mechanisms and functional roles

  • Explore specialized functions in different tissues and organisms

Environmental Influences on Glutarylation:

• Methodological approach:

  • Expose cells/organisms to environmental stressors

  • Quantify glutarylation changes using the antibody

  • Correlate with metabolic alterations and transcriptional responses

  • Investigate transgenerational inheritance of glutarylation patterns

Integration with Structural Biology:

• Methodological approach:

  • Use antibodies to purify glutarylated histones for structural studies

  • Apply cryo-EM to visualize glutarylated nucleosomes

  • Perform molecular dynamics simulations to predict structural effects

  • Develop structure-based approaches to targeting glutarylation

These emerging research directions highlight the potential of Glutaryl-HIST1H2BC (K116) Antibody to contribute to fundamental advances in our understanding of epigenetic regulation and its implications for human health and disease .

What is the current consensus on the biological significance of HIST1H2BC glutarylation in different cellular contexts and disease states?

The current consensus regarding HIST1H2BC glutarylation is still evolving, but several key insights have emerged from recent research:

Fundamental Biological Roles:

• Transcriptional regulation:

  • Glutarylation at K116 appears to correlate with transcriptional activation in many contexts

  • The modification likely alters chromatin compaction by neutralizing the positive charge of lysine

  • Evidence suggests glutarylation may function in recruiting specific transcriptional machinery

• Metabolic sensing:

  • HIST1H2BC glutarylation serves as a link between cellular metabolism and gene expression

  • Levels fluctuate in response to changes in TCA cycle intermediates and glutaryl-CoA availability

  • May represent a mechanism for adapting transcriptional programs to metabolic states

• Cell cycle regulation:

  • Emerging evidence suggests dynamic changes in glutarylation during cell cycle progression

  • May play roles in regulating replication timing and chromosomal segregation

  • Often works in concert with other histone modifications in a coordinated manner

Disease Associations:

• Cancer biology:

  • Altered glutarylation patterns have been observed in several cancer types

  • May contribute to oncogenic transcriptional programs and metabolic rewiring

  • Potential biomarker value for certain cancer subtypes or stages

• Metabolic disorders:

  • Dysregulated glutarylation in conditions with altered energy metabolism

  • Potential involvement in insulin resistance and diabetes pathophysiology

  • Links to mitochondrial dysfunction in metabolic syndrome

• Neurodegenerative diseases:

  • Preliminary evidence of altered histone glutarylation in models of neurodegeneration

  • May influence neuronal gene expression patterns and stress responses

  • Potential therapeutic target for maintaining neural homeostasis

Regulatory Mechanisms:

• Enzymatic control:

  • "Writers": Several acyltransferases can catalyze glutarylation, though specificity remains unclear

  • "Erasers": Sirtuin family members (particularly SIRT5) remove glutaryl groups

  • "Readers": Proteins that specifically recognize glutarylated histones are being identified

• Cross-regulation with other modifications:

  • Glutarylation may compete with acetylation, methylation at shared lysine residues

  • Evidence for sequential modification patterns during cellular responses

  • Coordination with non-histone protein glutarylation in signaling networks

Current Research Gaps:

• Limited temporal and spatial resolution of glutarylation dynamics in living cells

• Incomplete understanding of site-specific functions (K116 vs. other sites)

• Need for more disease-specific studies to establish causal relationships

• Technical challenges in distinguishing closely related acyl modifications