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Product name
Anti-Histone H4 antibody
See all Histone H4 products (11) ...
Description
Rabbit polyclonal to Histone H4 - ChIP Grade
Tested applications
IP, ChIP, IHC-P, ICC/IF, WBmore details
Cross reactivity
Reacts with
Mouse, Rat, Human, Saccharomyces cerevisiae, Xenopus laevis, Fruit fly (Drosophila melanogaster)
Predicted to work with
Chicken, Cow, Pig, Caenorhabditis elegans
Immunogen
Positive control
This antibody gave a positive control in the following lysates: Histone prep HeLa Histone This antibody gave a positive control in the following mouse lysates: NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell MEF1 (Mouse embryonic fibroblast cell line) Whole Cell This antibody gave a positive control in the following rat lysate: PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Applications
Show applications key
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.
IP: Use at an assay dependent concentration.
ChIP: Use 2 µg for 25 µg of chromatin.
IHC-P: Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF: 1/200.
WB: Use a concentration of 1 µg/ml. Detects a band of approximately 14 kDa (predicted molecular weight: 11 kDa).Can be blocked with Histone H4 peptide (ab13843).
Target
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
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.
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.
Target information above from: UniProt accessionP62805
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- dJ160A22.1 antibody
- dJ160A22.2 antibody
- dJ221C16.1 antibody
see all
Database links
- Entrez Gene: 318846 Fruit fly (Drosophila melanogaster)
- Entrez Gene: 3771854 Fruit fly (Drosophila melanogaster)
- Entrez Gene: 3771893 Fruit fly (Drosophila melanogaster)
- Entrez Gene: 3771908 Fruit fly (Drosophila melanogaster)
- Entrez Gene: 3771935 Fruit fly (Drosophila melanogaster)
- Entrez Gene: 3771938 Fruit fly (Drosophila melanogaster)
- Entrez Gene: 3772509 Fruit fly (Drosophila melanogaster)
- Entrez Gene: 3772519 Fruit fly (Drosophila melanogaster)
see all
Anti-Histone H4 antibody images:
Western blot - Histone H4 antibody (ab10158)
All lanes : Anti-Histone H4 antibody (ab10158) at 1 µg/ml
Lane 1 : Histone prep
Lane 2 : HeLa histone lysate
Lane 3 : Histone prep with
Lane 4 : HeLa Histone lysate with
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 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?)
Western blot - Histone H4 antibody (ab10158)
All lanes : Anti-Histone H4 antibody (ab10158) at 1 µg/ml
Lane 1 :
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
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?)
ChIP - Anti-Histone H4 antibody (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.
Immunoprecipitation - Anti-Histone H4 antibody (ab10158)
Histone H4 was immunoprecipitated using 0.5mg NIH3T3 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H4 antibody (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.
References for Anti-Histone H4 antibody (ab10158)
This product has been referenced in:
- Bowman A et al. The histone chaperones nap1 and vps75 bind histones h3 and h4 in a tetrameric conformation. Mol Cell 41:398-408 (2011).Read more (PubMed: 21329878) »
- Hainer SJ & Martens JA Identification of histone mutants that are defective for transcription-coupled nucleosome occupancy. Mol Cell Biol 31:3557-68 (2011).Read more (PubMed: 21730290) »
See all 21 publications for this product
Publishing research using ab10158? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"