Acetyl-HIST1H3A (K115) Antibody
Description
Definition and Overview of Acetyl-HIST1H3A (K115) Antibody
The Acetyl-HIST1H3A (K115) Antibody is a polyclonal or recombinant antibody designed to detect acetylation at lysine 115 (K115) of histone H3.1, a variant of the histone H3 family. Acetylation at this site is an epigenetic modification linked to chromatin remodeling, transcriptional activation, and cellular processes such as differentiation and stress response .
| Parameter | Details |
|---|---|
| Target | Acetylated lysine 115 on HIST1H3A (Histone H3.1) |
| Host | Rabbit (polyclonal) or recombinant monoclonal (varies by vendor) |
| Immunogen | Synthetic peptide containing Acetyl-K115 from human HIST1H3A |
| Reactivity | Human (validated); predicted for other species (e.g., canine) |
| Applications | Western blotting (WB), immunocytochemistry (ICC), ELISA, immunofluorescence (IF) |
| Product Promise | Covered by vendor guarantees (e.g., Abcam, Biomatik) |
Western Blotting (WB)
Abcam’s ab240888 shows robust signal in lysates from human cell lines (HeLa, HEK-293, A549, K562) treated with sodium butyrate, with untreated controls serving as negative controls .
| Cell Line | Treatment | Band Intensity | Control |
|---|---|---|---|
| HeLa | + Sodium butyrate | Strong (15 kDa) | Untreated (-) |
| HEK-293 | + Sodium butyrate | Strong (15 kDa) | Untreated (-) |
| A549 | + Sodium butyrate | Moderate (15 kDa) | Untreated (-) |
Immunocytochemistry (ICC)
Biomatik’s CAC12903 and CAC12821 are validated for ICC, with protocols involving fixation, permeabilization, and biotinylated secondary detection .
ELISA and IF/ICC
Abbexa’s antibody is tested for ELISA and IF/ICC, though specific optimization details are not disclosed .
Challenges and Considerations
While no direct cross-reactivity data is reported for K115 acetyl antibodies, general pitfalls in histone PTM antibodies include:
Comparative Analysis of Available Antibodies
Future Directions and Recommendations
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Validation via Peptide Microarrays: Assess antibody specificity for K115 acetylation using platforms like those described in Rothbart et al. .
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ChIP-Seq Integration: Validate genomic binding patterns in cell lines with known acetylation profiles (e.g., sodium butyrate-treated vs. untreated).
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Cross-reactivity Testing: Screen against acetylated peptides at nearby lysines (e.g., K114, K116) to exclude off-target binding .
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Studies indicate that epigenetic regulation in cancer can occur by inducing E3 ubiquitin ligase NEDD4-dependent histone H3 ubiquitination. PMID: 28300060
- The identification of increased expression of H3K27me3 during a patient's clinical course can be useful for determining whether tumors are heterochronous. PMID: 29482987
- Research has demonstrated that JMJD5, a Jumonji C (JmjC) domain-containing protein, acts as a Cathepsin L-type protease that mediates histone H3 N-tail proteolytic cleavage under stress conditions that trigger a DNA damage response. PMID: 28982940
- Evidence suggests that the Ki-67 antigen proliferative index has significant limitations, and phosphohistone H3 (PHH3) is a viable alternative proliferative marker. PMID: 29040195
- These findings identify cytokine-induced histone 3 lysine 27 trimethylation as a mechanism that stabilizes gene silencing in macrophages. PMID: 27653678
- This data indicates that, in the early developing human brain, HIST1H3B constitutes the largest proportion of H3.1 transcripts among H3.1 isoforms. PMID: 27251074
- In a series of 47 diffuse midline gliomas, histone H3-K27M mutation was found to be mutually exclusive with IDH1-R132H mutation and EGFR amplification, rarely co-occurred with BRAF-V600E mutation, and was commonly associated with p53 overexpression, ATRX loss, and monosomy 10. Among these K27M+ diffuse midline gliomas. PMID: 26517431
- Data show that histone chaperone HIRA co-localizes with viral genomes, binds to incoming viral and deposits histone H3.3 onto these. PMID: 28981850
- Experiments have shown that PHF13 binds specifically to DNA and to two types of histone H3 methyl tags (lysine 4-tri-methyl or lysine 4-di-methyl) where it functions as a transcriptional co-regulator. PMID: 27223324
- Hemi-methylated CpGs DNA recognition activates UHRF1 ubiquitylation towards multiple lysines on the H3 tail adjacent to the UHRF1 histone-binding site. PMID: 27595565
- For the first time, this study describes the MR imaging features of pediatric diffuse midline gliomas with histone H3 K27M mutation. PMID: 28183840
- Approximately 30% of pediatric high grade gliomas (pedHGG) including GBM and DIPG harbor a lysine 27 mutation (K27M) in histone 3.3 (H3.3) which is correlated with poor outcome and was shown to influence EZH2 function. PMID: 27135271
- H3F3A K27M mutation in adult cerebellar HGG is not uncommon. PMID: 28547652
- Data show that lysyl oxidase-like 2 (LOXL2) is a histone modifier enzyme that removes trimethylated lysine 4 (K4) in histone H3 (H3K4me3) through an amino-oxidase reaction. PMID: 27735137
- Histone H3 lysine 9 (H3K9) acetylation was most prevalent when the Dbf4 transcription level was highest whereas the H3K9me3 level was greatest during and just after replication. PMID: 27341472
- SPOP-containing complex regulates SETD2 stability and H3K36me3-coupled alternative splicing. PMID: 27614073
- Data suggest that binding of the helical tail of histone 3 (H3) with PHD ('plant homeodomain') fingers of BAZ2A or BAZ2B (bromodomain adjacent to zinc finger domain 2A or 2B) requires molecular recognition of secondary structure motifs within the H3 tail and could represent an additional layer of regulation in epigenetic processes. PMID: 28341809
- The results demonstrate a novel mechanism by which Kdm4d regulates DNA replication by reducing the H3K9me3 level to facilitate formation of the preinitiation complex. PMID: 27679476
- Histone H3 modifications caused by traffic-derived airborne particulate matter exposures in leukocytes. PMID: 27918982
- A key role of persistent histone H3 serine 10 or serine 28 phosphorylation in chemical carcinogenesis through regulating gene transcription of DNA damage response genes. PMID: 27996159
- hTERT promoter mutations are frequent in medulloblastoma and are associated with older patients, prone to recurrence and located in the right cerebellar hemisphere. On the other hand, histone 3 mutations do not seem to be present in medulloblastoma. PMID: 27694758
- AS1eRNA-driven DNA looping and activating histone modifications promote the expression of DHRS4-AS1 to economically control the DHRS4 gene cluster. PMID: 26864944
- Data suggest that nuclear antigen Sp100C is a multifaceted histone H3 methylation and phosphorylation sensor. PMID: 27129259
- The authors propose that histone H3 threonine 118 phosphorylation via Aurora-A alters the chromatin structure during specific phases of mitosis to promote timely condensin I and cohesin disassociation, which is essential for effective chromosome segregation. PMID: 26878753
- Hemi-methylated DNA opens a closed conformation of UHRF1 to facilitate its H3 histone recognition. PMID: 27045799
- Functional importance of H3K9me3 in hypoxia, apoptosis and repression of APAK. PMID: 25961932
- Taken together, the authors verified that histone H3 is a real substrate for GzmA in vivo in the Raji cells treated by staurosporin. PMID: 26032366
- We conclude that circulating H3 levels correlate with mortality in sepsis patients and inversely correlate with antithrombin levels and platelet counts. PMID: 26232351
- Data show that double mutations on the residues in the interface (L325A/D328A) decreases the histone H3 H3K4me2/3 demethylation activity of lysine (K)-specific demethylase 5B (KDM5B). PMID: 24952722
- Data indicate that minichromosome maintenance protein 2 (MCM2) binding is not required for incorporation of histone H3.1-H4 into chromatin but is important for stability of H3.1-H4. PMID: 26167883
- Data suggest that histone H3 lysine methylation (H3K4me3) serves a crucial mechanistic role in leukemia stem cell (LSC) maintenance. PMID: 26190263
- PIP5K1A modulates ribosomal RNA gene silencing through its interaction with histone H3 lysine 9 trimethylation and heterochromatin protein HP1-alpha. PMID: 26157143
- Data indicate that lower-resolution mass spectrometry instruments can be utilized for histone post-translational modifications (PTMs) analysis. PMID: 25325711
- Data indicate that inhibition of lysine-specific demethylase 1 activity prevented IL-1beta-induced histone H3 lysine 9 (H3K9) demethylation at the microsomal prostaglandin E synthase 1 (mPGES-1) promoter. PMID: 24886859
- The authors report that de novo CENP-A assembly and kinetochore formation on human centromeric alphoid DNA arrays is regulated by a histone H3K9 acetyl/methyl balance. PMID: 22473132
Q&A
Basic Research Questions
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What is Acetyl-HIST1H3A (K115) and why is it significant in epigenetic research?
Acetyl-HIST1H3A (K115) refers to histone H3.1 protein that has been acetylated at lysine 115. Histone H3.1 is a core component of nucleosomes, which wrap and compact DNA into chromatin. Nucleosomes consist of 147 base pairs of DNA wrapped around an octamer of core histone proteins (two each of H2A, H2B, H3, and H4) .
This specific acetylation represents one of many post-translational modifications that constitute the "histone code" and plays crucial roles in regulating DNA accessibility. Core histone proteins are central to transcription regulation, DNA repair, DNA replication, and chromosomal stability . Acetylation neutralizes the positive charge of lysine residues, potentially weakening histone-DNA interactions and creating a more accessible chromatin structure.
Studying specific acetylation patterns at K115 provides valuable insights into how this particular modification contributes to gene regulation and chromatin organization within specific genomic contexts. This knowledge enhances our understanding of both normal cellular processes and disease mechanisms involving epigenetic dysregulation.
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What applications are appropriate for Acetyl-HIST1H3A (K115) antibodies?
Acetyl-HIST1H3A (K115) antibodies can be utilized in various experimental applications:
Application Recommended Dilution Reference Western Blotting (WB) 1:100-1:1000 Immunocytochemistry (ICC) 1:1-1:10 ELISA Varies by kit These antibodies can be used to detect and quantify the presence of Acetyl-HIST1H3A (K115) in cell lysates through Western blotting, visualize its cellular distribution through immunocytochemistry, and perform quantitative analysis through ELISA .
While not specifically mentioned for K115 antibodies in the search results, chromatin immunoprecipitation (ChIP) represents another common application for histone modification antibodies. ChIP allows researchers to identify genomic regions associated with specific histone modifications, and can be combined with DNA microarrays or next-generation sequencing for genome-wide analyses . This technique has revolutionized our ability to study the distribution of histone modifications across the genome.
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How do I verify the specificity of an Acetyl-HIST1H3A (K115) antibody?
Antibody specificity is crucial for reliable experimental results. Several approaches can verify specificity of Acetyl-HIST1H3A (K115) antibodies:
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Peptide competition assay: Incubate the antibody with a synthetic peptide containing acetylated K115 before experimentation. If the antibody is specific, the peptide should block binding to the target epitope, resulting in signal reduction.
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Histone mutant screening: Test the antibody against histones with mutations at K115 to confirm it doesn't bind when the target site is altered. As noted in search result , "ELISA does not assure antibody specificity in chromatin immunoprecipitation," making additional validation through histone mutations essential.
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Multiple antibody approach: Use several antibodies targeting the same modification but recognizing different epitopes to corroborate findings.
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Western blotting: Confirm the antibody detects a band of appropriate size (approximately 15-17 kDa for histone H3.1) .
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Mass spectrometry validation: Confirm acetylation at K115 in immunoprecipitated samples through mass spectrometry analysis.
Rigorous validation is particularly important for epigenetic studies, as cross-reactivity with other histone modifications can lead to misinterpretation of results and irreproducible findings.
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What are the best protocols for Western blotting with Acetyl-HIST1H3A (K115) antibodies?
Based on protocols used for similar histone acetylation antibodies, the following recommendations apply for Western blotting with Acetyl-HIST1H3A (K115) antibodies:
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Sample preparation: Extract histones carefully using acid extraction or commercial histone extraction kits. For whole cell lysates, 30 μg of protein per lane is typically sufficient .
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Gel preparation: Use a 12-15% SDS-PAGE gel to achieve good separation of histones (15-17 kDa) .
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Electrophoresis conditions: Run at 80V through stacking gel, then 120V through resolving gel for approximately 2 hours .
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Transfer: Transfer proteins to nitrocellulose membrane at 150 mA for 50-90 minutes .
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Blocking: Block membrane with 5% non-fat milk in TBS for 1.5 hours at room temperature .
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Primary antibody: Dilute Acetyl-HIST1H3A (K115) antibody 1:100-1:1000 in blocking buffer and incubate overnight at 4°C .
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Washing: Wash membrane with TBS-0.1% Tween, 3 times for 5 minutes each .
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Secondary antibody: Use appropriate HRP-conjugated secondary antibody (typically anti-rabbit IgG if primary antibody is rabbit-derived) .
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Detection: Develop signal using ECL substrate and appropriate imaging system .
Optimization may be necessary for your specific experimental system, as antibody performance can vary between lots and manufacturers.
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How does acetylation at K115 compare to other histone H3 acetylation sites?
While the search results don't provide specific information about functional differences between K115 acetylation and other sites, some inferences can be made from related research:
Acetylation of histone H3.1 at lysine 14 (K14) is known to function in "transcriptional activation, chromatin accessibility, cellular identity, epigenetic memory, and coordinated gene regulation" . Different acetylation sites often have distinct functional roles and may be regulated by specific enzymes.
A study using highly specific antibodies against various histone acetylation sites in yeast found that "telomeric and silent mating locus heterochromatin is hypoacetylated at all histone sites" . Additionally, different histone acetyltransferases and deacetylases affected specific acetylation sites differently. For example, "RPD3 is required for strongly deacetylating all sites except H4 K16, ESA1 for acetylating H2A, H2B, and H4 sites except H4 K16, and GCN5 for acetylating H2B and H3 sites except H3 K14" .
To determine the specific functional importance of K115 acetylation, comparative studies using antibodies targeting different acetylation sites, along with functional assays to assess impacts on transcription and chromatin structure, would be necessary.
