Product nameAnti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9]
See all Histone H4 primary antibodies
DescriptionMouse monoclonal [6F8-D9] to Histone H4 (di methyl K20, tri methyl K20)
Tested applicationsSuitable for: IHC-P, WB, ELISA, Dot blot, ICC/IF, Flow Cytmore details
Species reactivityReacts with: Mouse, Human, Caenorhabditis elegans
Predicted to work with: Xenopus laevis, Arabidopsis thaliana, Drosophila melanogaster, Mammals
Synthetic peptide within Human Histone H4 (tri methyl K20). The exact sequence is proprietary.
- ICC/IF: HeLa cell line
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
Constituents: PBS, 6.97% L-Arginine
Concentration information loading...
Light chain typekappa
Our Abpromise guarantee covers the use of ab78517 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 11 kDa.|
|ELISA||Use at an assay dependent concentration.|
|Dot blot||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 5 µg/ml.|
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
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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ab78517 stained in HeLa cells. Cells were fixed with 4% paraformaldehyde (10 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% Triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab78517 at 5µg/ml and ab6046 (Rabbit polyclonal to beta Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.
All lanes : Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9] (ab78517) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate
Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H4 peptide (ab14963) at 0.5 µg/ml
Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate with
Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µg/ml
Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate with
Human Histone H3 (di methyl K4) peptide (ab7768) at 0.5 µg/ml
Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate with
Human Histone H3 (tri methyl K4) peptide (ab1342) at 0.5 µg/ml
Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate with
Human Histone H4 (mono methyl K20) peptide (ab17043) at 0.5 µg/ml
Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H4 peptide (17-24) - di methyl K20 at 0.5 µg/ml
Lane 8 : Calf Thymus Histone Preparation Nuclear Lysate with
Human Histone H4 (tri methyl K20) peptide (ab17567) at 0.5 µg/ml
Lane 9 : Calf Thymus Histone Preparation Nuclear Lysate with
Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µg/ml
Lane 10 : Calf Thymus Histone Preparation Nuclear Lysate with
Human Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg/ml
Lane 11 : Calf Thymus Histone Preparation Nuclear Lysate with
Human Histone H3 (tri methyl K9) peptide (ab1773) at 0.5 µg/ml
Lysates/proteins at 0.5 µg per lane.
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 11 kDa
Observed band size: 13 kDa why is the actual band size different from the predicted?
Exposure time: 90 seconds
ab78517 is specific for Histone H4 (tri-methyl K20). This is illustrated in lane 8 where the activity of ab78517 is specifically blocked by the addition of the immunizing peptide (ab17567).
ICC/IF image of ab78517 stained mouse oocyte. Oocytes were fixed in formaldehyde and then incubated in 3% serum with 0.25% Triton for 30 minutes at 37°C. Cells were incubated with the antibody (ab78517) at 1/200 for 2 hours at 37°C. The secondary antibody used was a polyclonal Goat-anti-mouse conjugated to Alexa Fluor® 488 (1/250).
IHC image of Histone H4 (tri methyl K20) staining in normal human skin formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab78517, 1µg/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.
Overlay histogram showing HeLa cells stained with ab78517 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab78517, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed.
This product has been referenced in:
- Flora P et al. Transient transcriptional silencing alters the cell cycle to promote germline stem cell differentiation in Drosophila. Dev Biol 434:84-95 (2018). WB . Read more (PubMed: 29198563) »
- Ma Y & Buttitta L Chromatin organization changes during the establishment and maintenance of the postmitotic state. Epigenetics Chromatin 10:53 (2017). IHC . Read more (PubMed: 29126440) »