Recombinant Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184] (ab177309)

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.
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.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18184
Isotype

IgG
Research areas

Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

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
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.

Notes
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.

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
modifications

Acetylation 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.
Target information above from: UniProt accession P62805 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
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
- Entrez Gene: 8359 Human
- Entrez Gene: 8360 Human
- Entrez Gene: 8361 Human
- Entrez Gene: 8362 Human
- Entrez Gene: 8363 Human
- Entrez Gene: 8364 Human
- Entrez Gene: 8365 Human
- Entrez Gene: 8366 Human
- Entrez Gene: 8367 Human
- Entrez Gene: 8368 Human
- Entrez Gene: 8370 Human
- Entrez Gene: 100041230 Mouse
- Entrez Gene: 102641229 Mouse
- Entrez Gene: 319155 Mouse
- Entrez Gene: 319156 Mouse
- Entrez Gene: 319157 Mouse
- Entrez Gene: 319158 Mouse
- Entrez Gene: 319159 Mouse
- Entrez Gene: 319160 Mouse
- Entrez Gene: 319161 Mouse
- Entrez Gene: 319166 Mouse
- Entrez Gene: 320332 Mouse
- Entrez Gene: 326619 Mouse
- Entrez Gene: 326620 Mouse
- Entrez Gene: 69386 Mouse
- Entrez Gene: 97122 Mouse
- Entrez Gene: 64646 Rat
- GenBank: NM_003548 Human
- Omim: 142720 Human
- Omim: 142750 Human
- Omim: 602786 Human
- Omim: 602797 Human
- SwissProt: P04908 Human
- SwissProt: P62805 Human
- SwissProt: Q93077 Human
- SwissProt: Q99878 Human
- SwissProt: P10812 Mouse
- SwissProt: P22752 Mouse
- SwissProt: P62806 Mouse
- SwissProt: P02262 Rat
- Unigene: 121017 Human
- Unigene: 143080 Human
- Unigene: 247816 Human
- Unigene: 248172 Human
- Unigene: 248174 Human
- Unigene: 248178 Human
- Unigene: 248179 Human
- Unigene: 278483 Human
- Unigene: 352191 Human
- Unigene: 408067 Human
- Unigene: 417332 Human
- Unigene: 434195 Human
- Unigene: 46423 Human
- Unigene: 528055 Human
- Unigene: 533295 Human
- Unigene: 55468 Human
- Unigene: 591790 Human
- Unigene: 655235 Human
- Unigene: 662174 Human
- Unigene: 706635 Human
- Unigene: 745457 Human
- Unigene: 14775 Mouse
- Unigene: 158272 Mouse
- Unigene: 227295 Mouse
- Unigene: 228709 Mouse
- Unigene: 246720 Mouse
- Unigene: 255646 Mouse
- Unigene: 260530 Mouse
- Unigene: 261642 Mouse
- Unigene: 261662 Mouse
- Unigene: 261664 Mouse
- Unigene: 261665 Mouse
- Unigene: 442307 Mouse
- Unigene: 486099 Mouse
- Unigene: 98465 Rat
see all
Alternative names
- H4_HUMAN antibody
- Hist4h4 antibody
- Histone H2A antibody
- Histone H4 antibody

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 seconds

Blocking/Dilution buffer: 5% BSA/TBST.
Dot Blot - Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184] (ab177309)

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.
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.
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.
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.
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.
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.
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.
Anti-Histone H2A (phospho S1) + Histone H4 (phospho S1) antibody [EPR18184] (ab177309)

Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

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Datasheet download

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Publishing research using ab177309? Please let us know so that we can cite the reference in this datasheet.

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

Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Compatible Secondaries

Isotype control

Applications

The Abpromise guarantee

Target

Function

Involvement in disease

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (1)

Customer reviews and Q&As

Post-translational
modifications