Recombinant Anti-Histone H4 antibody [EPR16599] - ChIP Grade (ab177840)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16599] to Histone H4 - ChIP Grade
- Suitable for: PepArr, ChIP, IHC-P, WB, ICC/IF
- Reacts with: Mouse, Rat, Human, Drosophila melanogaster, Recombinant fragment
Related conjugates and formulations
Overview
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Product name
Anti-Histone H4 antibody [EPR16599] - ChIP Grade
See all Histone H4 primary antibodies -
Description
Rabbit monoclonal [EPR16599] to Histone H4 - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: PepArr, ChIP, IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human, Drosophila melanogaster, Recombinant fragment
Predicted to work with: a wide range of other species -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa and NIH/3T3 whole cell lysates; Drosophila whole lysate. ICC/IF: HeLa cells. IHC-P: Human colon, mouse pancreas and rat cerebral cortex tissues. ChIP: Chromatin from 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, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR16599 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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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
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab177840 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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ChIP |
Use 2 µg for 25 µg of chromatin.
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB | (4) |
1/1000. Detects a band of approximately 11 kDa (predicted molecular weight: 11 kDa).
We recommend using 3% milk as the blocking agent for Western blot. |
ICC/IF |
1/100.
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Notes |
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PepArr
Use at an assay dependent concentration. |
ChIP
Use 2 µg for 25 µg of chromatin. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Detects a band of approximately 11 kDa (predicted molecular weight: 11 kDa). We recommend using 3% milk as the blocking agent for Western blot. |
ICC/IF
1/100. |
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 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.
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. -
Cellular localization
Nucleus. Chromosome. - Information by UniProt
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Database links
- Entrez Gene: 318846 Drosophila melanogaster
- Entrez Gene: 3771854 Drosophila melanogaster
- Entrez Gene: 3771893 Drosophila melanogaster
- Entrez Gene: 3771908 Drosophila melanogaster
- Entrez Gene: 3771935 Drosophila melanogaster
- Entrez Gene: 3771938 Drosophila melanogaster
- Entrez Gene: 3771941 Drosophila melanogaster
- Entrez Gene: 3771947 Drosophila melanogaster
see all -
Alternative names
- dJ160A22.1 antibody
- dJ160A22.2 antibody
- dJ221C16.1 antibody
see all
Images
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Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with 0.75% formaldehyde for 10min. The ChIP was performed with 25μg of chromatin, 2μg of ab177840 (blue), and 20μl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (SYBR approach). Primers are located in the first kb of the transcribed region.
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All lanes : Anti-Histone H4 antibody [EPR16599] - ChIP Grade (ab177840) at 1/1000 dilution
Lanes 1 & 6 : Histone H1 Recombinant Protein
Lanes 2 & 7 : Histone H2A Recombinant Protein
Lanes 3 & 8 : Histone H2B Recombinant Protein
Lanes 4 & 9 : Histone H3.1 Recombinant Protein
Lanes 5 & 10 : Histone H4 Recombinant Protein
Lysates/proteins at 0.1 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (H+L), Peroxidase Conjugated at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 11 kDa
Observed band size: 15 kDa why is the actual band size different from the predicted?
Exposure time: 4 minutesLanes 1-5: 1% BSA blocking buffer
Lanes 6-10: 3% Milk blocking buffer
We recommend using 3% milk as the blocking agent for Western blot.
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Anti-Histone H4 antibody [EPR16599] - ChIP Grade (ab177840) at 1/1000 dilution + Drosophila whole lysates at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 11 kDa
Observed band size: 11 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Histone H4 antibody [EPR16599] - ChIP Grade (ab177840) at 1/5000 dilution
Lane 1 : HeLa whole cell lysates
Lane 2 : NIH/3T3 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.
Performed under reducing conditions.
Predicted band size: 11 kDa
Observed band size: 11 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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ab177840 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. -
Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 antibody [EPR16599] - ChIP Grade (ab177840)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling Histone H4 with ab177840 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Nuclear staining on HeLa cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
1. ab177840 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 antibody [EPR16599] - ChIP Grade (ab177840)
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histone H4 with ab177840 at 1/2000 dilution, followed by prediluted Goat Anti-Rabbit IgG H&L (HRP). Nucleus staining on glandular epithelium of Human colon tissue is observed. Counter stained with Hematoxylin.
Negative control: PBS instead of primary antibody; secondary antibody is prediluted 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 H4 antibody [EPR16599] - ChIP Grade (ab177840)
Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling Histone H4 with ab177840 at 1/2000 dilution, followed by prediluted Goat Anti-Rabbit IgG H&L (HRP). Nucleus staining on glandular epithelium of mouse pancreas tissue is observed. Counter stained with Hematoxylin.
Negative control: PBS instead of primary antibody; secondary antibody is prediluted 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 H4 antibody [EPR16599] - ChIP Grade (ab177840)
Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex tissue labeling Histone H4 with ab177840 at 1/2000 dilution, followed by prediluted Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining on neuron cells of cerebral cortex tissue is observed. Counter stained with Hematoxylin.
Negative control: PBS instead of primary antibody; secondary antibody is prediluted 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 (15)
ab177840 has been referenced in 15 publications.
- Xue X et al. Tumour cells are sensitised to ferroptosis via RB1CC1-mediated transcriptional reprogramming. Clin Transl Med 12:e747 (2022). PubMed: 35220675
- Shen T et al. An enhancer variant at 16q22.1 predisposes to hepatocellular carcinoma via regulating PRMT7 expression. Nat Commun 13:1232 (2022). PubMed: 35264579
- Merges GE et al. Loss of Prm1 leads to defective chromatin protamination, impaired PRM2 processing, reduced sperm motility and subfertility in male mice. Development 149:N/A (2022). PubMed: 35608054
- Fei D et al. KAT6A regulates stemness of aging bone marrow-derived mesenchymal stem cells through Nrf2/ARE signaling pathway. Stem Cell Res Ther 12:104 (2021). PubMed: 33541408
- Alanazi S et al. Tryptase Regulates the Epigenetic Modification of Core Histones in Mast Cell Leukemia Cells. Front Immunol 12:804408 (2021). PubMed: 34925389