Anti-Histone H4 (acetylated) (acetyl K8) antibody - ChIP Grade (ab15823)
Overview
- Product nameAnti-Histone H4 (acetylated) (acetyl K8) antibody - ChIP GradeSee all Histone H4 (acetylated) primary antibodies ...
- DescriptionRabbit polyclonal to Histone H4 (acetylated) (acetyl K8) - ChIP Grade
- Tested applicationsICC/IF, IHC-P, Electron Microscopy, ChIP, WB more details
- Species reactivityReacts with: Mouse, Cow, Human, Saccharomyces cerevisiae, Schizosaccharomyces pombe
- Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H4, acetylated at K8.
(Peptide available as ab15824.)
- Positive controlThis antibody gave a positive signal on Hela Histone lysate-Butyrate treated and Calf Thymus Histone lysate.
Properties
- FormLiquid
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS. pH 7.4 -
Concentration information loading... - PurityImmunogen affinity purified
- Clonality Polyclonal
- IsotypeIgG
- Research Areas
Applications
Our Abpromise guarantee covers the use of ab15823 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| ICC/IF | ICC/IF: Use a concentration of 1 µg/ml. |
| IHC-P | IHC-P: Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
| Electron Microscopy | EM: Use at an assay dependent dilution. PubMed: 20543957 |
| ChIP | ChIP: Use 2µl for 106 cells. |
| WB | WB: Use a concentration of 1 µg/ml. Detects a band of approximately 12 kDa (predicted molecular weight: 11.2 kDa).Can be blocked with Histone H4 peptide - acetyl K8 (ab15824). |
Target
- FunctionCore 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 similaritiesBelongs 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 localizationNucleus. Chromosome.
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Database links
- Entrez Gene: 280691 Cow
- Entrez Gene: 121504 Human
- Entrez Gene: 554313 Human
- Entrez Gene: 8294 Human
- Entrez Gene: 8359 Human
- Entrez Gene: 8360 Human
- Entrez Gene: 8361 Human
- Entrez Gene: 8368 Human
- Entrez Gene: 8370 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: 100041230 Mouse
- Entrez Gene: 319155 Mouse
- Entrez Gene: 319156 Mouse
- Entrez Gene: 319157 Mouse
- Entrez Gene: 319158 Mouse
- Entrez Gene: 319159 Mouse
- Entrez Gene: 69386 Mouse
- Entrez Gene: 97122 Mouse
- Entrez Gene: 319160 Mouse
- Entrez Gene: 319161 Mouse
- Entrez Gene: 320332 Mouse
- Entrez Gene: 326619 Mouse
- Entrez Gene: 326620 Mouse
- Entrez Gene: 674678 Mouse
- Omim: 142750 Human
- SwissProt: P62805 Human
- SwissProt: P62806 Mouse
- SwissProt: P02309 Saccharomyces cerevisiae
- SwissProt: P09322 Schizosaccharomyces pombe
- Unigene: 113 Cow
- Unigene: 352191 Human
- Unigene: 143080 Human
- Unigene: 247816 Human
- Unigene: 248172 Human
- Unigene: 248178 Human
- Unigene: 248179 Human
- Unigene: 278483 Human
- Unigene: 662174 Human
- Unigene: 706635 Human
- Unigene: 718935 Human
- Unigene: 46423 Human
- Unigene: 528055 Human
- Unigene: 533295 Human
- Unigene: 55468 Human
- Unigene: 591790 Human
- Unigene: 655235 Human
- Unigene: 144300 Mouse
- Unigene: 14775 Mouse
- Unigene: 158272 Mouse
- Unigene: 227295 Mouse
- Unigene: 228709 Mouse
- Unigene: 246720 Mouse
- Unigene: 442307 Mouse
- Unigene: 255646 Mouse
- Unigene: 260530 Mouse
- Unigene: 261642 Mouse
- Unigene: 261662 Mouse
- Unigene: 261664 Mouse
- Unigene: 377875 Mouse
see all
Target information above from: UniProt accession
P62805
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
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Alternative names
- H4 antibodyH4 antibodyH4_HUMAN antibody
- H4F2 antibodyH4F2 antibodyHIST4H4 antibodyHIST4H4 antibodyHistone H4 antibody
see all
Anti-Histone H4 (acetylated) (acetyl K8) antibody - ChIP Grade images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H4 (acetyl K8) antibody - ChIP Grade (ab15823)IHC image of Histone H4 (acetyl K8) 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 ab15823, 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. -
ChIP - Anti-Histone H4 (acetylated) (acetyl K8) antibody - ChIP Grade (ab15823)Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 µg of chromatin, 2 µg of ab15823 (blue), and 20 µl 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). Primers and probes are located in the first kb of the transcribed region.
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Western blot - Histone H4 (acetyl K8) antibody - ChIP Grade (ab15823)All lanes : Anti-Histone H4 (acetylated) (acetyl K8) antibody - ChIP Grade (ab15823) at 1 µg/ml
Lane 1 : Sodium butyrate treated hela histone
Lane 2 : Untreated hela histone
Lane 3 : Sodium butyrate treated hela histone withHistone H4 peptide - acetyl K8 (ab15824) at 1 µg/ml
Lane 4 : Untreated hela histone withHistone H4 peptide - acetyl K8 (ab15824) at 1 µg/ml
Lane 5 : Sodium butyrate treated hela histone withHistone H4 peptide (ab15825) at 1 µg/ml
Lane 6 : Untreated hela histone withHistone H4 peptide (ab15825) at 1 µg/ml
Lane 7 : Sodium butyrate treated hela histone with Histone H4 peptide - acetyl K5 (ab15596) at 1 µg/ml
Lane 8 : Untreated hela histone with Histone H4 peptide - acetyl K5 (ab15596) at 1 µg/ml
Lane 9 : Sodium butyrate treated hela histone withHistone H4 peptide - acetyl K12 (ab15662) at 1 µg/ml
Lane 10 : Untreated hela histone withHistone H4 peptide - acetyl K12 (ab15662) at 1 µg/ml
Lane 11 : Sodium butyrate treated hela histone with Histone H2A peptide - acetyl K9 at 1 µg/ml
Lane 12 : Untreated hela histone with Histone H2A peptide - acetyl K9 at 1 µg/ml
Lysates/proteins at 2.5 µg per lane.
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 11.2 kDa
Observed band size : 12 kDa (why is the actual band size different from the predicted?)
Additional bands at : 22 kDa (possible cross reactivity). -
Immunocytochemistry/ Immunofluorescence - Histone H4 (acetyl K8) antibody - ChIP Grade (ab15823)ICC/IF image of ab15823 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab15823, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). -
Electron Microscopy - Histone H4 (acetyl K8) antibody - ChIP Grade (ab15823)Image from Kizilyaprak C et al, PLoS One. 2010 Jun 9;5(6):e11039, Fig 5.A: Overall view of a murine rod photoreceptor nucleus immunogold labelled with acetylated H4K8-specific antibodies.B: Schematic representation of the H4K8ac-specific labelling. The bar represents 1µm. -
Western blot - Histone H4 (acetyl K8) antibody - ChIP Grade (ab15823)All lanes : Anti-Histone H4 (acetylated) (acetyl K8) antibody - ChIP Grade (ab15823) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate (ab121)
Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) withHistone H4 peptide (ab15825) at 0.5 µg/ml
Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) withHistone H4 peptide - acetyl K8 (ab15824) at 0.5 µg/ml
Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Histone H4 peptide - acetyl K5 at 0.5 µg/ml
Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) withHistone H4 peptide - acetyl K12 (ab15662) at 0.5 µg/ml
Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) withHistone H2A peptide - acetyl K9 (ab54016) at 0.5 µg/ml
Lysates/proteins at 0.5 µg/ml per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 11.2 kDa
Observed band size : 13 kDa (why is the actual band size different from the predicted?)
Exposure time : 3 minutes
References for Anti-Histone H4 (acetylated) (acetyl K8) antibody - ChIP Grade (ab15823)
This product has been referenced in:
- Schwartz BE et al. Differentiation of NUT midline carcinoma by epigenomic reprogramming. Cancer Res 71:2686-96 (2011). Read more (PubMed: 21447744) »
- Goossens E et al. Mouse germ cells go through typical epigenetic modifications after intratesticular tissue grafting. Hum Reprod 26:3388-400 (2011). Read more (PubMed: 21984576) »
