SUV39H2 Antibody

What is SUV39H2 and what cellular functions does it perform?

SUV39H2 (Suppressor of Variegation 3-9 Homolog 2) is a histone methyltransferase that specifically trimethylates lysine 9 of histone H3 (H3K9), using monomethylated H3K9 as its substrate . This enzyme plays a critical role in epigenetic transcriptional repression by creating H3K9 trimethylation marks that recruit heterochromatin protein 1 (HP1) family proteins. SUV39H2 functions predominantly in heterochromatin regions, playing a central role in establishing constitutive heterochromatin at pericentric and telomere regions . During embryogenesis, SUV39H2 overlaps in tissue expression with SUV39H1, but in adult organisms, SUV39H2 transcripts become restricted to the testes . The protein shows enrichment at heterochromatin from the leptotene to round spermatid stage during spermatogenesis, with specific accumulation on sex chromosome chromatin during the first meiotic prophase .

What are the typical applications for SUV39H2 antibodies in research?

SUV39H2 antibodies are utilized in multiple experimental techniques including:

For optimal results, researchers should titrate the antibody concentration in each testing system as effectiveness can be sample-dependent . Multiple publications have validated these applications, with Western blot being the most commonly cited technique across human and mouse samples .

How should SUV39H2 antibodies be stored and handled to maintain reactivity?

Proper storage is crucial for maintaining antibody performance:

• Store at -20°C for long-term preservation

• Aliquot the antibody before freezing to avoid repeated freeze-thaw cycles that can degrade protein structure and function

• Most formulations are stable for one year after shipment when properly stored

• For antibodies in liquid form, they typically come in PBS buffer containing preservatives such as sodium azide (0.02-0.09%) and may contain stabilizers like glycerol (up to 50%)

When working with the antibody, thaw aliquots completely before use and keep on ice during experimental procedures. Some preparations contain 0.1% BSA in small volume formats (20 µL) for additional stability .

What species reactivity can be expected with commercial SUV39H2 antibodies?

The reactivity profile varies by antibody preparation:

When selecting an antibody for cross-species applications, it's advisable to review sequence homology data and published validation studies for the specific antibody clone. The UniProt ID for human SUV39H2 is Q9H5I1, which can be used to compare sequence conservation across species of interest .

How can I differentiate between SUV39H1 and SUV39H2 in experimental systems?

Distinguishing between SUV39H1 and SUV39H2 requires careful experimental design due to their 59% sequence identity in mice :

• Antibody selection: Use highly specific antibodies raised against unique peptide regions. For example, antibodies targeting the C-terminal region (amino acids 367-395) of human SUV39H2 can provide specificity .

• Expression pattern analysis: SUV39H2 transcripts become restricted to testes in adult animals, while SUV39H1 shows broader expression. This tissue-specific pattern can help differentiate the proteins in adult tissues .

• Knockout/knockdown validation: Validate antibody specificity using genetic models where either SUV39H1 or SUV39H2 is depleted. Several publications have used knockdown approaches to confirm antibody specificity .

• Molecular weight distinction: SUV39H2 has a calculated molecular weight of approximately 47-53 kDa, which may differ slightly from SUV39H1 on Western blots, allowing for differentiation when running both proteins on the same gel .

• Functional assays: During spermatogenesis, SUV39H2 specifically accumulates with chromatin of sex chromosomes during meiotic silencing, providing a functional context for differentiation .

What are the best practices for validating SUV39H2 antibody specificity?

• Knockout/knockdown controls: The gold standard for antibody validation is testing in systems where the target protein has been genetically deleted or depleted. Published literature has used knockdown approaches to validate SUV39H2 antibodies .

• Blocking peptide competition: Pre-incubate the antibody with the immunizing peptide before application to samples. Signal disappearance confirms specificity to the target epitope.

• Multiple antibody concordance: Use antibodies from different sources or raised against different epitopes of SUV39H2. Consistent results strengthen confidence in specificity.

• Recombinant protein controls: Include purified or overexpressed SUV39H2 as a positive control in Western blot experiments to confirm correct molecular weight detection (approximately 47-53 kDa) .

• Cross-reactivity testing: Test the antibody against related proteins (particularly SUV39H1) to ensure it doesn't cross-react with similar proteins in the methyltransferase family.

How can SUV39H2 antibodies be optimized for chromatin immunoprecipitation (ChIP) experiments?

Although not explicitly listed in the tested applications from our search results, researchers interested in ChIP applications should consider:

• Crosslinking optimization: As SUV39H2 interacts with heterochromatin regions, optimizing formaldehyde crosslinking time (typically 10-15 minutes) is crucial for capturing these interactions without overfixing.

• Sonication parameters: Heterochromatin regions can be more resistant to sonication. Extend sonication time or increase power to ensure proper fragmentation of heterochromatic regions where SUV39H2 operates.

• Antibody selection: Choose antibodies that have been affinity-purified and validated for nuclear proteins, such as those purified through protein A columns followed by peptide affinity purification .

• Input controls: Due to the enrichment of SUV39H2 at specific genomic regions like pericentric heterochromatin, careful normalization to input controls is essential.

• Validation with known targets: Include primers for known SUV39H2-enriched regions, such as pericentric repeats or telomeric regions, as positive controls in ChIP-qPCR validation.

What special considerations exist for detecting SUV39H2 in testicular tissues?

Since SUV39H2 shows testis-specific expression in adult animals and plays important roles during spermatogenesis , researchers should consider:

• Sample preparation: Testicular tissues contain diverse cell populations at different stages of spermatogenesis. Consider using techniques like laser capture microdissection to isolate specific cell types (leptotene to round spermatid stages) where SUV39H2 is enriched.

• Fixation protocols: Optimize fixation methods for testicular tissues, which can be challenging due to the compact nature of seminiferous tubules. Perfusion fixation may provide better results than immersion fixation.

• Stage-specific analysis: Design experiments to capture the dynamic localization of SUV39H2, which accumulates at sex chromosome chromatin during meiotic silencing in the first meiotic prophase .

• Co-staining strategies: Use markers for specific spermatogenic stages alongside SUV39H2 antibodies to correlate expression with developmental timing.

• Signal amplification: Consider using signal amplification methods if the endogenous levels of SUV39H2 are difficult to detect in tissue sections.

How can researchers investigate the role of SUV39H2 in pathological conditions?

Recent publications have implicated SUV39H2 in several pathological processes :

• Nonalcoholic steatohepatitis: Studies have shown that the histone methyltransferase contributes to this condition in mice, suggesting examination of liver tissues with appropriate controls .

• Intervertebral disc degeneration: Research has demonstrated that SUV39H2-mediated lysine methylation of PPP1CA disrupts TFEB-dependent autophagy, promoting intervertebral disc degeneration .

• Viral infection models: Studies have examined how SUV39H2 interacts with the Epstein-Barr virus polymerase processivity factor at the intersection of transcription and replication .

When investigating these conditions:

• Use appropriate tissue or cell models relevant to the condition

• Include disease and healthy control samples

• Consider temporal dynamics of SUV39H2 expression during disease progression

• Combine protein expression analysis with functional assays of H3K9 trimethylation

• Validate findings with genetic manipulation of SUV39H2 levels

What are common issues when using SUV39H2 antibodies in Western blotting?

When troubleshooting Western blot experiments with SUV39H2 antibodies:

• Molecular weight confirmation: The calculated molecular weight of SUV39H2 is approximately 47-53 kDa . Bands at significantly different sizes may indicate non-specific binding or protein degradation.

• Recommended dilutions: Start with the manufacturer's recommended dilution (typically 1:500-1:2000 for Western blotting) and optimize as needed for your specific sample type .

• Positive controls: HEK-293 cells have been validated as positive Western blot controls for some SUV39H2 antibodies .

• Sample preparation: Nuclear proteins like SUV39H2 require effective nuclear extraction protocols. Ensure your lysis buffer effectively solubilizes nuclear proteins while preserving epitope integrity.

• Transfer conditions: For higher molecular weight proteins, extend transfer time or adjust buffer conditions to ensure complete transfer to the membrane.

How can epigenetic inheritance studies benefit from SUV39H2 antibodies?

SUV39H2 plays potential roles in epigenetic imprinting in the male germline :

• Developmental timing analysis: Track SUV39H2 localization throughout spermatogenesis using antibodies validated for immunohistochemistry or immunofluorescence.

• Inheritance tracking: Combine SUV39H2 immunostaining with H3K9me3 detection to correlate the enzyme's presence with its epigenetic mark across generations.

• Experimental models: Utilize conditional knockout models with temporally controlled SUV39H2 deletion to examine the precise windows when its activity affects epigenetic inheritance.

• Chromatin state analysis: Pair SUV39H2 antibody studies with techniques like ATAC-seq to correlate the enzyme's presence with changes in chromatin accessibility.

• Sperm epigenome profiling: Use SUV39H2 antibodies in ChIP-seq experiments on purified sperm cells to map potential heritable H3K9me3 patterns established by this enzyme.