Anti-Histone H2A (phospho S129) antibody (ab15083)
Key features and details
- Rabbit polyclonal to Histone H2A (phospho S129)
- Suitable for: WB, ELISA, PepArr
- Reacts with: Schizosaccharomyces pombe
- Isotype: IgG
Overview
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Product name
Anti-Histone H2A (phospho S129) antibody
See all Histone H2A primary antibodies -
Description
Rabbit polyclonal to Histone H2A (phospho S129) -
Host species
Rabbit -
Tested applications
Suitable for: WB, ELISA, PepArrmore details -
Species reactivity
Reacts with: Schizosaccharomyces pombe
Predicted to work with: Saccharomyces cerevisiaeDoes not react with: Human -
Immunogen
Synthetic peptide corresponding to Saccharomyces cerevisiae Histone H2A aa 100 to the C-terminus (phospho S129) conjugated to bovine serum albumin.
Database link: P04911
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentrationConcentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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ChIP Related Products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab15083 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application Abreviews Notes WB 1/500 - 1/1000. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa). ELISA Use at an assay dependent concentration. PepArr Use a concentration of 0.2 - 0.02 µg/ml. Target
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Function
Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. -
Sequence similarities
Belongs to the histone H2A family. -
Post-translational
modificationsThe chromatin-associated form is phosphorylated on Thr-121 during mitosis.
Deiminated on Arg-4 in granulocytes upon calcium entry.
Monoubiquitination of Lys-120 by RING1 and RNF2/RING2 complex gives a specific tag for epigenetic transcriptional repression and participates in X chromosome inactivation of female mammals. It is involved in the initiation of both imprinted and random X inactivation. Ubiquitinated H2A is enriched in inactive X chromosome chromatin. Ubiquitination of H2A functions downstream of methylation of 'Lys-27' of histone H3. Monoubiquitination of Lys-120 by RNF2/RING2 can also be induced by ultraviolet and may be involved in DNA repair. Following DNA double-strand breaks (DSBs), it is ubiquitinated through 'Lys-63' linkage of ubiquitin moieties by the E2 ligase UBE2N and the E3 ligases RNF8 and RNF168, leading to the recruitment of repair proteins to sites of DNA damage. Monoubiquitination and ionizing radiation-induced 'Lys-63'-linked ubiquitination are distinct events.
Phosphorylation on Ser-2 is enhanced during mitosis. Phosphorylation on Ser-2 by RPS6KA5/MSK1 directly represses transcription. Acetylation of H3 inhibits Ser-2 phosphorylation by RPS6KA5/MSK1.
Symmetric dimethylation on Arg-4 by the PRDM1/PRMT5 complex may play a crucial role in the germ-cell lineage. -
Cellular localization
Nucleus. Chromosome. - Information by UniProt
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Database links
- SwissProt: P04912 Saccharomyces cerevisiae
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Alternative names
- H2a 615 antibody
- H2A antibody
- H2A GL101 antibody
see all
Images
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All lanes : Anti-Histone H2A (phospho S129) antibody (ab15083) at 1/500 dilution
Lane 1 : S.cerevisiae yeast extract (SCYE) + control peptide with Human Histone H2A peptide (ab19751)
Lane 2 : S.cerevisiae yeast extract with 0.2 % Methyl methanesulfonate (1 hour) with Human Histone H2A peptide (ab19751)
Lane 3 : SCYE + phospho peptide with Histone H2A peptide - phospho S129 (ab19828)
Lane 4 : SCYE_M + phospho peptide with Histone H2A peptide - phospho S129 (ab19828)
Lysates/proteins at 10 µg per lane.
Performed under reducing conditions.
Predicted band size: 14 kDaThe blots were produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membranes were then blocked for an hour. 1 microgram per mL of control- (ab19751, lane 1 and 2) or phospho-peptides (ab19828, lane 3 and 4) were added to the primary antibody ab15083 (rabbit anti-Histone H2A (phospho S129) antibody; diluted 1:500) and loading control ab125247 (mouse anti-GAPDH antibody; diluted 1:20000) and the membranes were incubated with peptide/antibody mixture for 24 hours at 4°C. Antibody binding was detected using infrared (IR)-labelled goat anti-rabbit (green) and IR-labelled goat anti-mouse (red; insert below) antibodies, diluted 1:20,000, for 1 hour at room temperature before imaging.
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Anti-Histone H2A (phospho S129) antibody (ab15083) at 1 µg/ml + S.cerevisiae (Y190) Whole Cell Lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 14 kDa
Additional bands at: 17 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 90 seconds -
All batches of ab15083 are tested in Peptide Array against peptides to different Histone H2A modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H2A - phospho S129 (ab19828), indicating that this antibody specifically recognises the Histone H2A - phospho S129 modification.
ab19828 - Histone H2A - phospho S129
ab19751 - Histone H2A - unmodified
Datasheets and documents
References (72)
ab15083 has been referenced in 72 publications.
- Chong SY et al. H3K4 methylation at active genes mitigates transcription-replication conflicts during replication stress. Nat Commun 11:809 (2020). PubMed: 32041946
- Diernfellner ACR et al. A pathway linking translation stress to checkpoint kinase 2 signaling in Neurospora crassa. Proc Natl Acad Sci U S A 116:17271-17279 (2019). PubMed: 31413202
- Bhalla P et al. Yeast PAF1 complex counters the pol III accumulation and replication stress on the tRNA genes. Sci Rep 9:12892 (2019). PubMed: 31501524
- Morafraile EC et al. Checkpoint inhibition of origin firing prevents DNA topological stress. Genes Dev 33:1539-1554 (2019). PubMed: 31624083
- House NC et al. Distinct roles for S. cerevisiae H2A copies in recombination and repeat stability, with a role for H2A.1 threonine 126. Elife 8:N/A (2019). PubMed: 31804179
Images
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All lanes : Anti-Histone H2A (phospho S129) antibody (ab15083) at 1/500 dilution
Lane 1 : S.cerevisiae yeast extract (SCYE) + control peptide with Human Histone H2A peptide (ab19751)
Lane 2 : S.cerevisiae yeast extract with 0.2 % Methyl methanesulfonate (1 hour) with Human Histone H2A peptide (ab19751)
Lane 3 : SCYE + phospho peptide with Histone H2A peptide - phospho S129 (ab19828)
Lane 4 : SCYE_M + phospho peptide with Histone H2A peptide - phospho S129 (ab19828)
Lysates/proteins at 10 µg per lane.
Performed under reducing conditions.
Predicted band size: 14 kDaThe blots were produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membranes were then blocked for an hour. 1 microgram per mL of control- (ab19751, lane 1 and 2) or phospho-peptides (ab19828, lane 3 and 4) were added to the primary antibody ab15083 (rabbit anti-Histone H2A (phospho S129) antibody; diluted 1:500) and loading control ab125247 (mouse anti-GAPDH antibody; diluted 1:20000) and the membranes were incubated with peptide/antibody mixture for 24 hours at 4°C. Antibody binding was detected using infrared (IR)-labelled goat anti-rabbit (green) and IR-labelled goat anti-mouse (red; insert below) antibodies, diluted 1:20,000, for 1 hour at room temperature before imaging.
-
Anti-Histone H2A (phospho S129) antibody (ab15083) at 1 µg/ml + S.cerevisiae (Y190) Whole Cell Lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 14 kDa
Additional bands at: 17 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 90 seconds
-
All batches of ab15083 are tested in Peptide Array against peptides to different Histone H2A modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H2A - phospho S129 (ab19828), indicating that this antibody specifically recognises the Histone H2A - phospho S129 modification.
ab19828 - Histone H2A - phospho S129
ab19751 - Histone H2A - unmodified