Product nameAnti-Histone H4 (symmetric di methyl R3) antibody - ChIP Grade
See all Histone H4 primary antibodies
DescriptionRabbit polyclonal to Histone H4 (symmetric di methyl R3) - ChIP Grade
Tested applicationsSuitable for: IHC-P, PepArr, ChIP, ICC/IF, WBmore details
Species reactivityReacts with: Mouse, Cow, Human, Saccharomyces cerevisiae, Xenopus laevis, Arabidopsis thaliana, Drosophila melanogaster, Tobacco
Predicted to work with: Rat, Pig, Rainbow trout, Corn
Synthetic peptide within Histone H4 aa 1-100 (symmetric di methyl R3) conjugated to Keyhole Limpet Haemocyanin (KLH). The exact sequence is proprietary.
(Peptide available as
- WB: Calf thymus histone preparation and HeLa whole cell extract. IHC-P: Human normal kidney FFPE tissue sections.
Storage instructionsShipped 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 bufferpH: 7.40
Preservative: 0.02% Sodium azide
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...
PurityImmunogen affinity purified
ChIP Related Products
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab5823 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|PepArr||Use a concentration of 0.2 - 0.02 µg/ml.|
|ChIP||Use 2-4 µg for 25 µg of chromatin.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 14 kDa (predicted molecular weight: 11.5 kDa).Can be blocked with Human Histone H4 (symmetric di methyl R3) peptide (ab14791).|
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.
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.
- Information by UniProt
- 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
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Anti-Histone H4 (symmetric di methyl R3) antibody - ChIP Grade (ab5823) at 1 µg + Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg
Anti-Rabbit IgG VHH Single Domain (HRP) (ab191866) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 11.5 kDa
Observed band size: 14 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutes
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab5823 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 2µg of ab5823 (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.
Interphase HeLa cell immunofluorescently labelled with anti-dimethylArg3 (H2A + H4) antibody (ab5823, 1/2000) and a polyclonal goat anti-Rabbit secondary antibody conjugated to Cy3 (1/200). ab5823 shows a strong nuclear focal staining pattern. For more information please see Abreview.
IHC image of ab5823 staining Histone H4 (symmetric di methyl R3) in human kidney formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. 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 ab5823, 5µg/ml, for 15 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. No primary antibody was used in the negative control (shown on the inset).
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
All batches of ab5823 are tested in Peptide Array against peptides to different Histone H4 and H2A modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H4 - symmetric di methyl R3 peptide (ab14791), indicating that this antibody specifically recognises the Histone H4 - symmetric di methyl R3 modification.
ab14791 - Histone H4 - symmetric di methyl R3
ab17416 - Histone H4 asymmetric di methyl R3
ab15821 - Histone H4 - unmodified
ab22399 - Histone H2A symmetric di methyl R3
ab22398 - Histone H2A asymmetric di methyl R3
ab13186 - Histone H2A - unmodified
This product has been referenced in:
- Gao Y & Ge W The histone methyltransferase DOT1L inhibits osteoclastogenesis and protects against osteoporosis. Cell Death Dis 9:33 (2018). Read more (PubMed: 29348610) »
- Wei D et al. Transcriptional Regulation by CpG Sites Methylation in the Core Promoter Region of the Bovine SIX1 Gene: Roles of Histone H4 and E2F2. Int J Mol Sci 19:N/A (2018). Read more (PubMed: 29337851) »