Di-Methyl-Histone H4 (Lys79) Antibody
Product Specs
Target Background
- Studies demonstrate that PP32 and SET/TAF-Ibeta proteins inhibit HAT1-mediated H4 acetylation. PMID: 28977641
- Research suggests that post-translational modifications of histones, specifically trimethylation of lysine 36 in H3 (H3K36me3) and acetylation of lysine 16 in H4 (H4K16ac), play roles in DNA damage repair. H3K36me3 stimulates H4K16ac upon DNA double-strand breaks, and SETD2, LEDGF, and KAT5 are essential for these epigenetic changes. (SETD2 = SET domain containing 2; LEDGF = lens epithelium-derived growth factor; KAT5 = lysine acetyltransferase 5) PMID: 28546430
- Data indicate that Omomyc protein co-localizes with proto-oncogene protein c-myc (c-Myc), protein arginine methyltransferase 5 (PRMT5), and histone H4 H4R3me2s-enriched chromatin domains. PMID: 26563484
- H4K12ac is regulated by estrogen receptor-alpha and is associated with BRD4 function and inducible transcription. PMID: 25788266
- Systemic lupus erythematosus appears to be linked to an imbalance in histone acetyltransferases and histone deacetylase enzymes, favoring pathological H4 acetylation. PMID: 25611806
- Sumoylated human histone H4 prevents chromatin compaction by inhibiting long-range internucleosomal interactions. PMID: 25294883
- Acetylation at lysine 5 of histone H4 is associated with lytic gene promoters during reactivation of Kaposi's sarcoma-associated herpesvirus. PMID: 25283865
- An increase in histone H4 acetylation induced by hypoxia in human neuroblastoma cell lines corresponds to elevated levels of N-myc transcription factor in these cells. PMID: 24481548
- Data suggest that G1-phase histone assembly is restricted to CENP-A and H4. PMID: 23363600
- This study investigated the distribution of a specific histone modification, namely H4K12ac, in human sperm and characterized its specific enrichment sites in promoters throughout the human genome. PMID: 22894908
- SRP68/72 heterodimers function as major nuclear proteins whose binding of the histone H4 tail is inhibited by H4R3 methylation. PMID: 23048028
- TNF-alpha inhibition of AQP5 expression in human salivary gland acinar cells is attributed to an epigenetic mechanism involving suppression of acetylation of histone H4. PMID: 21973049
- Findings suggest that global histone H3 and H4 modification patterns serve as potential markers for tumor recurrence and disease-free survival in non-small cell lung cancer. PMID: 22360506
- HAT1 differentially impacts nucleosome assembly of H3.1-H4 and H3.3-H4. PMID: 22228774
- Phosphorylation of histone H4 Ser 47, catalyzed by the PAK2 kinase, promotes nucleosome assembly of H3.3-H4 and inhibits nucleosome assembly of H3.1-H4 by increasing the binding affinity of HIRA to H3.3-H4 and reducing the association of CAF-1 with H3.1-H4. PMID: 21724829
- The imatinib-induced hemoglobinization and erythroid differentiation in K562 cells are associated with global histone H4 modification. PMID: 20949922
- Research reveals the molecular mechanisms by which DNA sequences within specific gene bodies are sufficient to nucleate the monomethylation of histone H4 lysine 200, which, in turn, reduces gene expression by half. PMID: 20512922
- Histone H4 expression is downregulated by zinc and upregulated by docosahexaenoate in a neuroblastoma cell line. PMID: 19747413
- Low levels of histone acetylation are associated with the development and progression of gastric carcinomas, possibly through alteration of gene expression. PMID: 12385581
- Overexpression of MTA1 protein and acetylation levels of histone H4 protein are closely related. PMID: 15095300
- Peptidylarginine deiminase 4 regulates histone Arg methylation by converting methyl-Arg to citrulline and releasing methylamine. Data suggest that PAD4 mediates gene expression by regulating Arg methylation and citrullination in histones. PMID: 15345777
- The lack of biotinylation of K12 in histone H4 is an early signaling event in response to double-strand breaks. PMID: 16177192
- Incorporation of acetylated histone H4-K16 into nucleosomal arrays inhibits the formation of compact 30-nanometer-like fibers and impedes the ability of chromatin to form cross-fiber interactions. PMID: 16469925
- Apoptosis is associated with global DNA hypomethylation and histone deacetylation events in leukemia cells. PMID: 16531610
- BTG2 contributes to retinoic acid activity by favoring differentiation through a gene-specific modification of histone H4 arginine methylation and acetylation levels. PMID: 16782888
- A relationship exists between histone H4 modification, epigenetic regulation of BDNF gene expression, and long-term memory for extinction of conditioned fear. PMID: 17522015
- The H4 tail and its acetylation have novel roles in mediating the recruitment of multiple regulatory factors that can alter chromatin states for transcription regulation. PMID: 17548343
- Brd2 bromodomain 2 is monomeric in solution and dynamically interacts with H4-AcK12; additional secondary elements in the long ZA loop may be a common characteristic of BET bromodomains. PMID: 17848202
- Spermatids Hypac-H4 impairment in mixed atrophy did not deteriorate further by AZFc region deletion. PMID: 18001726
- The SET8 and PCNA interaction couples H4-K20 methylation with DNA replication. PMID: 18319261
- H4K20 monomethylation and PR-SET7 are crucial for L3MBTL1 function. PMID: 18408754
- High expression of acetylated H4 is more prevalent in aggressive than indolent cutaneous T-cell lymphoma. PMID: 18671804
- Findings indicate a significant role of histone H4 modifications in bronchial carcinogenesis. PMID: 18974389
- Results demonstrate that acetylation of histone H4 K16 during S-phase enables early replicating chromatin domains to acquire the H4K16ac-K20me2 epigenetic label, which persists on the chromatin throughout mitosis and is deacetylated in early G1-phase of the next cell cycle. PMID: 19348949
- Acetylated H4 is overexpressed in diffuse large B-cell lymphoma and peripheral T-cell lymphoma compared to normal lymphoid tissue. PMID: 19438744
- The release of histone H4 by holocrine secretion from the sebaceous gland may play a critical role in innate immunity. PMID: 19536143
- Histone modification, including PRC2-mediated repressive histone marker H3K27me3 and active histone marker acH4, may be involved in CD11b transcription during HL-60 leukemia cells reprogramming to terminal differentiation. PMID: 19578722
- A role of Cdk7 in regulating elongation is further suggested by enhanced histone H4 acetylation and diminished histone H4 trimethylation on lysine 36—two marks of elongation—within genes when the kinase was inhibited. PMID: 19667075
- Data revealed the dynamic fluctuation of histone H4 acetylation levels during mitosis, as well as acetylation changes in response to structurally distinct histone deacetylase inhibitors. PMID: 19805290
- Data directly implicate BBAP in the monoubiquitylation and additional posttranslational modification of histone H4 and an associated DNA damage response. PMID: 19818714
Q&A
Basic Research Questions
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What is Di-Methyl-Histone H4 (Lys79) and what is its biological significance?
Di-Methyl-Histone H4 (Lys79) refers to histone H4 that has been dimethylated at the lysine residue at position 79. Histones are basic nuclear proteins responsible for the nucleosome structure of chromosomal fiber in eukaryotes. The nucleosome consists of approximately 146 bp of DNA wrapped around an octamer comprised of pairs of the four core histones (H2A, H2B, H3, and H4) . Histone methylation is a critical epigenetic modification that regulates gene expression. Methylation events that weaken binding between histone tails and DNA typically lead to increased transcription by making DNA more accessible to transcription factor proteins and RNA polymerase . While H3K79 methylation has been extensively studied and shown to be associated with active transcription in coding regions , H4K79 dimethylation represents another important histone modification involved in chromatin regulation.
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How should I prepare samples for optimal results with Di-Methyl-Histone H4 (Lys79) Antibody?
For optimal results with Di-Methyl-Histone H4 (Lys79) Antibody, follow these methodological steps:
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Western Blotting preparation: Use a dilution of 1:500-1:1000
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Sample preparation: For histone extraction, use acid extraction methods that preserve post-translational modifications
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Storage conditions: Store antibody at -20°C and avoid repeated freeze-thaw cycles
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Buffer composition: The antibody is typically supplied in PBS with 0.02% sodium azide and 50% glycerol at pH 7.4
Application Recommended Dilution Validated Species Western Blotting 1:500-1:1000 Human, Mouse, Rat Immunoprecipitation Not specified Not specified ChIP Not specified Not specified -
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How specific is the Di-Methyl-Histone H4 (Lys79) Antibody?
Di-Methyl-Histone H4 (Lys79) Antibody is highly specific for the dimethylated lysine 79 of histone H4. Based on patterns observed with similar histone modification antibodies, these antibodies are typically validated for specificity using:
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Dot blot analysis against panels of modified histone peptides
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Western blotting against recombinant histones with defined modifications
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Immunoprecipitation followed by mass spectrometry
High-quality antibodies should not cross-react with:
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What are the storage and handling requirements for Di-Methyl-Histone H4 (Lys79) Antibody?
For optimal antibody performance and longevity:
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Store at -20°C in the dark
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Avoid repeated freeze-thaw cycles by preparing small working aliquots
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The antibody is typically supplied in PBS with 0.02% sodium azide and 50% glycerol pH 7.4
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When working with the antibody, keep it on ice
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Centrifuge the antibody vial before opening to ensure all liquid is at the bottom
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For long-term storage of diluted antibody solutions, add carrier proteins like BSA (0.1-1%) to prevent adsorption to surfaces
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Monitor antibody performance over time as sensitivity may decrease with extended storage
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Technical Applications and Methodologies
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What controls should be included when using Di-Methyl-Histone H4 (Lys79) Antibody for ChIP experiments?
For rigorous ChIP experiments with Di-Methyl-Histone H4 (Lys79) Antibody, include these essential controls:
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Experimental controls:
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Input DNA (pre-immunoprecipitation chromatin) - crucial for normalization
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IgG control (same species as the primary antibody) - measures non-specific binding
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No-antibody control - assesses background from beads/reagents
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Positive control regions - known to be enriched for the modification
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Negative control regions - known to lack the modification
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Antibody validation controls:
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Peptide competition assay - confirms specificity
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Immunoprecipitation followed by mass spectrometry - verifies target
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Dot blot against modified peptide arrays - tests cross-reactivity
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Biological controls:
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Samples with known alterations in H4K79 methylation
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Treatment with methyltransferase inhibitors
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Cell types with different expression levels of methyltransferases
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Based on practices established for H3K79me2 ChIP protocols , a systematic approach including positive control genes (EIF2S3, CCT5) and negative control regions (inactive genes like MYOD1 and satellite repeats) provides robust validation.
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How can Di-Methyl-Histone H4 (Lys79) Antibody be used to study the interplay between different histone modifications?
To study the interplay between histone modifications, researchers can employ several sophisticated approaches:
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Sequential ChIP (Re-ChIP):
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Perform ChIP with Di-Methyl-Histone H4 (Lys79) Antibody
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Elute the immunoprecipitated chromatin
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Perform a second ChIP with antibodies against other modifications
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This identifies regions with co-occurrence of both modifications
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Integrated genomic analysis:
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Perform separate ChIP-seq experiments for multiple modifications
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Use computational approaches to identify regions of overlap or mutual exclusion
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Correlate with transcriptomic data to assess functional consequences
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Mass spectrometry-based approaches:
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Immunoprecipitate nucleosomes with Di-Methyl-Histone H4 (Lys79) Antibody
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Analyze co-occurring modifications using mass spectrometry
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Quantify the relative abundance of different modification patterns
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Proximity ligation assays:
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Use antibodies against different modifications
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Generate signal only when modifications are in close proximity
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Provides spatial information about co-occurring modifications
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The complex interplay between H4K79 dimethylation and other modifications likely functions within the broader context of the histone code, influencing transcription factor recruitment, chromatin accessibility, and gene expression.
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