Anti-Histone H4 antibody - ChIP Grade (ab10158)
Key features and details
- Rabbit polyclonal to Histone H4 - ChIP Grade
- Suitable for: IP, ChIP, IHC-P, WB
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
- Long-term and scalable supply – powered by recombinant technology for fast production
- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-Histone H4 antibody - ChIP Grade
See all Histone H4 primary antibodies -
Description
Rabbit polyclonal to Histone H4 - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: IP, ChIP, IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Chicken, Cow, Pig, Saccharomyces cerevisiae, Xenopus laevis, Caenorhabditis elegans, Drosophila melanogaster
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab13843) -
General notes
For recombinant monoclonal Histone H4 antibody (ChIP grade) - please see ab213291 (IGX4696H).
For mouse monoclonal Histone H4 antibody (ChIP grade) - please see ab31830 (mAbcam 31830).
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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ChIP Related Products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
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Related Products
- Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)
- Human Histone H4 (symmetric di methyl R3) peptide (ab14791)
- Histone H4 Total Quantification Kit (Colorimetric) (ab156909)
- Human Histone H4 (tri methyl K20) peptide (ab17567)
- Histone H4 Modification Multiplex Assay Kit (Colorimetric) (ab185914)
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab10158 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 |
|---|---|---|
| IP |
Use at an assay dependent concentration.
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| ChIP | (2) |
Use 2 µg for 25 µg of chromatin.
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| IHC-P | (1) |
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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| WB | (11) |
Use a concentration of 1 µg/ml. Detects a band of approximately 14 kDa (predicted molecular weight: 11 kDa).Can be blocked with Human Histone H4 peptide (ab13843).
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| Notes |
|---|
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IP
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
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 14 kDa (predicted molecular weight: 11 kDa).Can be blocked with Human Histone H4 peptide (ab13843). |
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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ChIP - Anti-Histone H4 antibody - ChIP Grade (ab10158)
Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab10158 (blue), and 20µl of protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.
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Western blot - Anti-Histone H4 antibody - ChIP Grade (ab10158)All lanes : Anti-Histone H4 antibody - ChIP Grade (ab10158) at 1 µg/ml
Lane 1 : Histone prep
Lane 2 : HeLa histone lysate
Lane 3 : Histone prep withHuman Histone H4 peptide (ab13843) at 1 µg/ml
Lane 4 : HeLa Histone lysate withHuman Histone H4 peptide (ab13843) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 11 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 antibody - ChIP Grade (ab10158)IHC image of Histone H4 staining in human breast carcinoma FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab10158, 1µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. -
Immunoprecipitation - Anti-Histone H4 antibody - ChIP Grade (ab10158)Histone H4 was immunoprecipitated using 0.5mg NIH/3T3 whole cell lysate, 5µg of Rabbit polyclonal to Histone H4 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, NIH3T3 whole cell lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab10158.
Secondary: Clean blot (HRP conjugate) at 1/1000 dilution.
Band: 14kDa: Histone H4; non specific - 52 and 85kDa: We are unsure as to the identity of this extra band. -
Western blot - Anti-Histone H4 antibody - ChIP Grade (ab10158)All lanes : Anti-Histone H4 antibody - ChIP Grade (ab10158) at 1 µg/ml
Lane 1 :NIH/3T3 whole cell lysate (ab7179)
Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 11 kDa
Observed band size: 14 kDa why is the actual band size different from the predicted?
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (242)
ab10158 has been referenced in 242 publications.
- Barrett E et al. Uteroplacental Insufficiency with Hypoxia Upregulates Placental PPAR?-KMT5A Axis in the Rat. Reprod Sci 28:1476-1488 (2021). PubMed: 33398850
- Shalini V et al. Genome-wide occupancy reveals the localization of H1T2 (H1fnt) to repeat regions and a subset of transcriptionally active chromatin domains in rat spermatids. Epigenetics Chromatin 14:3 (2021). PubMed: 33407810
- Slaughter MJ et al. HDAC inhibition results in widespread alteration of the histone acetylation landscape and BRD4 targeting to gene bodies. Cell Rep 34:108638 (2021). PubMed: 33472068
- Koussa NC & Smith DJ Limiting DNA polymerase delta alters replication dynamics and leads to a dependence on checkpoint activation and recombination-mediated DNA repair. PLoS Genet 17:e1009322 (2021). PubMed: 33493195
- Stefos GC et al. aniFOUND: analysing the associated proteome and genomic landscape of the repaired nascent non-replicative chromatin. Nucleic Acids Res 49:e64 (2021). PubMed: 33693861