Recombinant Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184] (ab177309)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18184] to Histone H2A (phospho S1) + Histone H4 (phospho S1)
- Suitable for: PepArr, Dot blot, WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184]
See all Histone H2A (phospho S1) + Histone H4 (phospho S1) primary antibodies -
Description
Rabbit monoclonal [EPR18184] to Histone H2A (phospho S1) + Histone H4 (phospho S1) -
Host species
Rabbit -
Tested applications
Suitable for: PepArr, Dot blot, WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa treated with 1.5µg/ml Colcemid for 12 hours whole cell lysates. IHC-P: human colon, mouse stomach, rat colon, human cerebral cortex, mouse heart and rat brain tissues. ICC/IF: HeLa cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18184 -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab177309 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 |
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PepArr |
Use at an assay dependent concentration.
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Dot blot |
1/1000.
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WB |
1/500. Detects a band of approximately 11, 14 kDa (predicted molecular weight: 11, 14 kDa).
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IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/4000.
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Notes |
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PepArr
Use at an assay dependent concentration. |
Dot blot
1/1000. |
WB
1/500. Detects a band of approximately 11, 14 kDa (predicted molecular weight: 11, 14 kDa). |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/4000. |
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. -
Involvement in disease
Chromosomal aberrations involving HISTONE H4 is a cause of B-cell non-Hodgkin lymphomas (B-cell NHL). Translocation t(3;6)(q27;p21), with BCL6. -
Sequence similarities
Belongs to the histone H4 family. -
Post-translational
modificationsAcetylation at Lys-6 (H4K5ac), Lys-9 (H4K8ac), Lys-13 (H4K12ac) and Lys-17 (H4K16ac) occurs in coding regions of the genome but not in heterochromatin.
Citrullination at Arg-4 (H4R3ci) by PADI4 impairs methylation.
Monomethylation and asymmetric dimethylation at Arg-4 (H4R3me1 and H4R3me2a, respectively) by PRMT1 favors acetylation at Lys-9 (H4K8ac) and Lys-13 (H4K12ac). Demethylation is performed by JMJD6. Symmetric dimethylation on Arg-4 (H4R3me2s) by the PRDM1/PRMT5 complex may play a crucial role in the germ-cell lineage.
Monomethylated, dimethylated or trimethylated at Lys-21 (H4K20me1, H4K20me2, H4K20me3). Monomethylation is performed by SET8. Trimethylation is performed by SUV420H1 and SUV420H2 and induces gene silencing.
Phosphorylated by PAK2 at Ser-48 (H4S47ph). This phosphorylation increases the association of H3.3-H4 with the histone chaperone HIRA, thus promoting nucleosome assembly of H3.3-H4 and inhibiting nucleosome assembly of H3.1-H4.
Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins. Monoubiquitinated at Lys-92 of histone H4 (H4K91ub1) in response to DNA damage. The exact role of H4K91ub1 in DNA damage response is still unclear but it may function as a licensing signal for additional histone H4 post-translational modifications such as H4 Lys-21 methylation (H4K20me).
Sumoylated, which is associated with transcriptional repression.
Crotonylation (Kcr) is specifically present in male germ cells and marks testis-specific genes in post-meiotic cells, including X-linked genes that escape sex chromosome inactivation in haploid cells. Crotonylation marks active promoters and enhancers and confers resistance to transcriptional repressors. It is also associated with post-meiotically activated genes on autosomes. -
Cellular localization
Nucleus. Chromosome. - Information by UniProt
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Database links
- Entrez Gene: 121504 Human
- Entrez Gene: 3012 Human
- Entrez Gene: 317772 Human
- Entrez Gene: 554313 Human
- Entrez Gene: 8294 Human
- Entrez Gene: 8335 Human
- Entrez Gene: 8337 Human
- Entrez Gene: 8338 Human
see all -
Alternative names
- H4_HUMAN antibody
- Hist4h4 antibody
- Histone H2A antibody
- Histone H4 antibody
Images
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Western blot - Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184] (ab177309)All lanes : Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184] (ab177309) at 1/500 dilution
Lane 1 : Untreated HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) treated with 1.5µg/ml Colcemid for 12 hours whole cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 11, 14 kDa
Observed band size: 11,14 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking/Dilution buffer: 5% BSA/TBST.
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Dot blot analysis of Histone H2A (phospho S1) peptide (Lane 1), unmodified Histone H2A peptide (Lane 2), Histone H4 (phospho S1) peptide (Lane 3), and unmodified Histone H4 peptide (Lane 4), labeled using ab177309 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184] (ab177309)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Histone H2A (phospho S1) + Histone H4 (phospho S1) with ab177309 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab177309 at 1/2000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/500 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution. -
Peptide Array - Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184] (ab177309)
ab177309 was tested in Peptide array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate).
Circle area represents affinity between the antibody and a peptide: all antigen-containing peptides are displayed as red circles, all other peptides as blue circles. The affinity is calculated as area under curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.
The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, can be downloaded here. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184] (ab177309)
Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Histone H2A (phospho S1) + Histone H4 (phospho S1) with ab177309 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.
Nuclear staining on human colon tissue is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184] (ab177309)
Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling Histone H2A (phospho S1) + Histone H4 (phospho S1) with ab177309 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.
Nuclear staining on mouse stomach tissue is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184] (ab177309)
Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Histone H2A (phospho S1) + Histone H4 (phospho S1) with ab177309 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.
Nuclear staining on rat colon tissue is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184] (ab177309)
Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling Histone H2A (phospho S1) + Histone H4 (phospho S1) with ab177309 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.
Nuclear staining on neuron cells of human cerebral cortex is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184] (ab177309)
Immunohistochemical analysis of paraffin-embedded mouse heart tissue labeling Histone H2A (phospho S1) + Histone H4 (phospho S1) with ab177309 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.
Nuclear staining on mouse heart tissue is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184] (ab177309)
Immunohistochemical analysis of paraffin-embedded rat brain tissue labeling Histone H2A (phospho S1) + Histone H4 (phospho S1) with ab177309 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.
Nuclear staining on neuron cells of rat brain tissue is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab177309 has been referenced in 1 publication.
- Chen T et al. Ultrastructure of spermiogenesis and the distribution of spermatozoal nuclear histones in the Japanese mantis shrimp, Oratosquilla oratoria (Crustacea: Stomatopoda). J Morphol 280:1170-1184 (2019). PubMed: 31141207