suv39h1a Antibody

What is SUV39H1 and what are its primary functions in cellular biology?

SUV39H1 (Suppressor of Variegation 3-9 Homolog 1) is a histone lysine methyltransferase responsible for the trimethylation of histone 3 lysine 9 (H3K9). It plays a crucial role in regulating heterochromatin formation, which inhibits transcription through compacting chromatin structure . SUV39H1 is one of two homologues of the Drosophila Suppressor-of-variegation 3(9) protein (Su(var)3-9) and contains both a SET domain involved in methyltransferase activity and a chromodomain that may bind to lysine 9 methylation, contributing to the regional spreading of this modification . H3K9 methylation is a hallmark of heterochromatin, appearing in transcriptionally silenced regions such as centromeres, repetitive elements, and inactive genes .

Which applications are suitable for SUV39H1 antibodies in epigenetic research?

SUV39H1 antibodies are valuable tools for various epigenetic research applications, including:

• Chromatin Immunoprecipitation (ChIP): For identifying SUV39H1 binding sites across the genome

• Western Blotting (WB): For detecting and quantifying SUV39H1 protein expression

• Immunocytochemistry (ICC): For visualizing SUV39H1 localization within cells

• Immunoprecipitation (IP): For studying protein-protein interactions involving SUV39H1

• Proximity Ligation Assay (PLA): For detecting protein interactions in situ

When selecting an antibody for these applications, researchers should verify the validation status for their specific application and consider factors such as host species, isotype, and purification method.

How should researchers validate SUV39H1 antibodies for their specific experimental systems?

Validation of SUV39H1 antibodies should follow a systematic approach:

• Specificity testing: Perform Western blot analysis in systems with known SUV39H1 expression levels, including knockdown or knockout controls to confirm specificity

• Cross-reactivity assessment: Test the antibody in multiple model organisms if working across species

• Application-specific validation:

  • For ChIP applications: Verify enrichment at known SUV39H1 binding sites

  • For immunofluorescence: Include appropriate controls and compare with published localization patterns

  • For Western blots: Confirm the detection of bands at the expected molecular weight (~48 kDa)

• Batch-to-batch consistency: When receiving new lots, compare performance with previously validated lots

What are the optimal conditions for using SUV39H1 antibody in Western blotting?

For optimal Western blotting results with SUV39H1 antibody:

• Sample preparation:

  • Extract total protein using standard lysis buffers containing protease inhibitors

  • Separate proteins on 10% SDS-PAGE gel

  • Transfer to PVDF membrane

• Blocking and antibody incubation:

  • Block membranes with 5% skim milk

  • Incubate with primary anti-SUV39H1 antibody (such as Cell Signaling Technology #8729)

  • Use HRP-conjugated anti-rabbit IgG as secondary antibody

  • Detect using chemiluminescent substrate (like SuperSignal West Femto Maximum Sensitivity Substrate)

• Optimization parameters:

  • Primary antibody dilution: Typically 1:1000, but optimize based on specific antibody

  • Incubation time: Overnight at 4°C for primary antibody

  • Washing buffer: PBS or TBS with 0.1% Tween-20

Remember to include appropriate loading controls such as GAPDH and to provide all blots with clear membrane edges in supplementary materials .

What are the key considerations for using SUV39H1 antibody in ChIP assays?

When performing ChIP assays with SUV39H1 antibody:

• Crosslinking optimization:

  • Standard 1% formaldehyde for 10 minutes works for most histone modification studies

  • Dual crosslinking with additional DSG may improve results for challenging targets

• Sonication parameters:

  • Optimize sonication conditions to achieve chromatin fragments of 200-500 bp

  • Verify fragmentation by agarose gel electrophoresis

• Antibody selection and validation:

  • Choose ChIP-validated antibodies like the MG44 clone (RRID: AB_2793343)

  • Verify enrichment at known SUV39H1 binding sites

• Controls to include:

  • Input chromatin (pre-immunoprecipitation)

  • IgG negative control

  • Positive control loci (heterochromatic regions)

  • Negative control loci (active genes not regulated by SUV39H1)

• Data analysis:

  • Normalize to input DNA

  • Compare with IgG control

  • Analyze relative enrichment at target loci

For higher sensitivity, consider using a specialized ChIP kit like ChIP-IT High Sensitivity or magnetic bead-based ChIP-IT Express Kits .

How can researchers effectively use SUV39H1 antibody in cell migration assays?

Based on published methodologies, here's an optimized protocol for using SUV39H1 antibody in cell migration studies:

• Experimental setup:

  • Seed cells (e.g., Hep3B cells) into 6-well plates

  • Transfect with SUV39H1 overexpression plasmids or siRNAs (for knockdown) for 48 hours

  • Create a wound by scraping the cell monolayer with sterile 10 μL plastic pipette tips

  • Wash cells twice to remove detached cells

  • Add medium containing 1% FBS to minimize proliferation effects

• Monitoring and analysis:

  • Capture images at defined timepoints (0h, 24h, 48h)

  • Measure wound closure using ImageJ software

  • Calculate migration rate as percentage of wound closure

• SUV39H1 expression verification:

  • In parallel wells, collect samples for Western blot

  • Verify SUV39H1 expression/knockdown using validated antibodies

  • Correlate SUV39H1 levels with observed migration phenotypes

This approach has successfully demonstrated that SUV39H1 overexpression promotes migration of hepatoma cells, while knockdown inhibits migration .

How can SUV39H1 antibody be utilized to investigate the relationship between SUV39H1 and oxidative phosphorylation (OXPHOS)?

Recent research has identified a novel connection between SUV39H1 and the OXPHOS pathway in hepatocellular carcinoma. To investigate this relationship using SUV39H1 antibody:

• Expression correlation studies:

  • Manipulate SUV39H1 levels (overexpression/knockdown)

  • Use SUV39H1 antibody for Western blot validation

  • Perform RNA-seq to identify differential expression of OXPHOS genes

  • Validate key targets by qRT-PCR and protein-level analysis

• Functional validation experiments:

  • Measure ATP production in cells with altered SUV39H1 expression

  • Use OXPHOS inhibitors (Rotenone, Oligomycin) in combination with SUV39H1 overexpression

  • Assess effects on cellular proliferation and migration

• Mechanistic studies:

  • Perform ChIP-seq with SUV39H1 antibody to identify direct binding to OXPHOS gene promoters

  • Analyze H3K9me3 marks at relevant genomic loci

  • Investigate protein-protein interactions between SUV39H1 and mitochondrial regulators

This approach has revealed that SUV39H1 overexpression upregulates OXPHOS pathway-related genes (COX6A1, COX6B1, COX8A, UQCRB, UQCR10, UQCRH, and NDUFA1) and increases ATP production, suggesting that SUV39H1 promotes metabolic reprogramming in cancer cells .

What are the best practices for using SUV39H1 antibody in studies examining HBV-related hepatocellular carcinoma?

For researchers investigating the role of SUV39H1 in HBV-related hepatocellular carcinoma:

• Cellular models and experimental design:

  • Compare HBV-positive (HepG2.215) and HBV-negative (HepG2) hepatoma cell lines

  • Transfect HepG2 cells with HBV whole genome plasmid as an infection model

  • Use SUV39H1 antibody to quantify protein levels by Western blot

• SUV39H1 manipulation strategies:

  • Overexpression: Transfect with SUV39H1 expression plasmids

  • Knockdown: Use siRNA or shRNA targeting SUV39H1

  • Pharmacological inhibition: Apply Chaetocin (SUV39H1 inhibitor)

  • Validate all manipulations using SUV39H1 antibody

• Functional assays:

  • Proliferation: CCK-8 assay, colony formation assay

  • Migration: Wound healing assay

  • Connect functional outcomes to SUV39H1 levels

• Clinical sample analysis:

  • Compare serum SUV39H1 levels across healthy controls, chronic hepatitis B patients, and HBV-HCC patients

  • Correlate with clinical indicators (ALT, AST, AFP)

  • Assess diagnostic potential by ROC curve analysis

Research using this approach has demonstrated that SUV39H1 is upregulated by HBV infection and promotes proliferation and migration of hepatoma cells, suggesting its potential as a diagnostic biomarker for HBV-HCC .

How can researchers troubleshoot inconsistent results when using SUV39H1 antibody across different experimental systems?

When facing inconsistent results with SUV39H1 antibody:

• Antibody-specific factors:

  Issue	Troubleshooting Approach
  Batch variation	Compare lot numbers; request certificate of analysis
  Non-specific binding	Optimize blocking conditions; try different blocking agents
  Degradation	Check storage conditions; prepare fresh working dilutions
  Epitope masking	Try different fixation methods for ICC/IF applications

• Biological variables to consider:

  • Cell type-specific expression patterns of SUV39H1

  • Influence of cell cycle phase on SUV39H1 levels

  • Post-translational modifications affecting antibody recognition

  • Splice variants present in different systems

• Technical optimization:

  • Adjust antibody concentration through titration experiments

  • Modify incubation times and temperatures

  • Evaluate different detection systems

  • Consider using multiple antibodies targeting different epitopes

• Validation strategies:

  • Include positive and negative control samples

  • Perform specificity tests using SUV39H1 knockdown/knockout samples

  • Compare results with alternative detection methods

  • Verify findings using orthogonal techniques

How can SUV39H1 antibody be used to investigate its potential as a cancer biomarker?

Recent studies suggest SUV39H1 has potential as a biomarker for various cancers, particularly HBV-associated hepatocellular carcinoma. To investigate this potential:

• Clinical sample analysis:

  • Compare serum SUV39H1 levels across patient cohorts using ELISA with validated antibodies

  • Analyze tissue expression by immunohistochemistry in tumor vs. adjacent normal tissue

  • Correlate SUV39H1 levels with clinical outcomes (survival, recurrence)

• Diagnostic performance assessment:

  • Calculate sensitivity, specificity, and area under ROC curve

  • Compare with established biomarkers (e.g., AFP for HCC)

  • Evaluate combined biomarker panels (SUV39H1 + AFP)

• Methodological considerations:

  • Standardize sample collection and processing

  • Establish reliable cutoff values

  • Validate findings in independent cohorts

Research has shown that serum SUV39H1 levels are higher in chronic hepatitis B patients than in healthy controls and higher in HBV-HCC patients than in CHB patients. When combined with AFP, SUV39H1 showed improved diagnostic value compared to AFP alone for HCC detection .

What are the considerations when using SUV39H1 antibody in multiplex immunofluorescence studies?

For researchers planning multiplex immunofluorescence studies with SUV39H1 antibody:

• Antibody panel design:

  • Select antibodies raised in different host species to avoid cross-reactivity

  • Consider antibody isotypes and secondary antibody specificity

  • Include markers for relevant cellular compartments (nuclear, mitochondrial)

  • Plan sequence of antibody application carefully

• Technical optimization:

  • Test each antibody individually before multiplexing

  • Optimize fixation and antigen retrieval conditions

  • Determine optimal antibody concentration for each marker

  • Establish appropriate blocking to minimize non-specific binding

• Controls and validation:

  • Single-color controls to assess bleed-through

  • Absorption controls to confirm specificity

  • Secondary-only controls to detect non-specific binding

  • Known positive and negative samples

• Analysis considerations:

  • Use appropriate spectral unmixing if needed

  • Establish quantitative analysis workflows

  • Consider colocalization metrics for interaction studies

  • Apply machine learning approaches for complex pattern recognition

How can ChIP-seq with SUV39H1 antibody be integrated with other omics approaches to understand epigenetic regulation?

An integrated multi-omics approach using SUV39H1 antibody can provide comprehensive insights into epigenetic regulation mechanisms:

• Experimental design for multi-omics integration:

  • Perform ChIP-seq with SUV39H1 antibody and H3K9me3 antibody

  • Combine with RNA-seq to correlate binding with gene expression

  • Include ATAC-seq to assess chromatin accessibility

  • Consider Hi-C for three-dimensional chromatin organization

  • Add DNA methylation analysis for complete epigenetic profiling

• Bioinformatic integration strategies:

  Approach	Application
  Peak overlap analysis	Identify genomic regions with multiple epigenetic marks
  Correlation analysis	Connect SUV39H1 binding with expression changes
  Network analysis	Construct gene regulatory networks
  Motif enrichment	Identify co-factors and binding motifs
  Machine learning	Predict functional outcomes from epigenetic patterns

• Validation experiments:

  • Confirm key findings with ChIP-qPCR using SUV39H1 antibody

  • Perform genetic perturbation of SUV39H1 followed by multi-omics

  • Use CUT&RUN or CUT&Tag for higher resolution binding profiles

This integrated approach has been applied to investigate SUV39H1's role in regulating the oxidative phosphorylation pathway in hepatocellular carcinoma, revealing its impact on metabolic reprogramming in cancer cells .

What emerging technologies are enhancing the utility of SUV39H1 antibody in epigenetic research?

Several cutting-edge technologies are expanding the applications of SUV39H1 antibody in epigenetic research:

• Advanced chromatin profiling methods:

  • CUT&RUN and CUT&Tag techniques provide higher resolution and lower background than traditional ChIP

  • ChIC (Chromatin Immunocleavage) offers improved sensitivity for limited sample inputs

  • Single-cell ChIP-seq enables analysis of epigenetic heterogeneity

• Spatial and temporal approaches:

  • Live-cell imaging with fluorescently tagged antibody fragments

  • Proximity labeling methods (BioID, APEX) to identify context-specific interaction partners

  • Optogenetic control of SUV39H1 activity combined with antibody-based detection

• Therapeutic and diagnostic applications:

  • Development of antibody-drug conjugates targeting SUV39H1 in cancer cells

  • SUV39H1-based liquid biopsy approaches for cancer detection

  • Companion diagnostics to predict response to epigenetic therapies

These technological advances promise to deepen our understanding of SUV39H1 function in health and disease, potentially leading to new diagnostic and therapeutic approaches.

How can researchers contribute to standardizing SUV39H1 antibody validation criteria for reproducible epigenetic research?

To improve reproducibility in SUV39H1 research:

• Comprehensive antibody validation:

  • Follow guidelines from the International Working Group for Antibody Validation

  • Perform genetic, orthogonal, independent antibody, and expression validation

  • Document detailed validation methods in publications

  • Share validation data through repositories and antibody validation databases

• Standardized reporting:

  • Provide complete antibody information (supplier, catalog number, lot, RRID)

  • Describe detailed experimental protocols

  • Share raw data and analysis code

  • Report negative results and limitations

• Community engagement:

  • Participate in antibody validation initiatives

  • Contribute to community standards for epigenetic research

  • Engage with organizations promoting research reproducibility

  • Share experiences through method-focused publications