Product nameAnti-Histone H4 (di methyl K20) antibody - ChIP Grade
See all Histone H4 primary antibodies
DescriptionRabbit polyclonal to Histone H4 (di methyl K20) - ChIP Grade
Tested applicationsSuitable for: IP, WB, ChIP, ICC/IF, IHC-Pmore details
Species reactivityReacts with: Mouse, Cow, Human, Schizosaccharomyces pombe, Toxoplasma gondii
Predicted to work with: Rat, a wide range of other speciesDoes not react with: Saccharomyces cerevisiae
Synthetic peptide within Saccharomyces cerevisiae Histone H4 aa 1-100 (di methyl K20) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
(Peptide available as
- WB: Calf thymus histone preparation and HeLa whole cell extract. IHC-P: FFPE human pancreas adenocarcinoma tissue. IF/ICC: 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 bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
ChIP Related Products
Our Abpromise guarantee covers the use of ab9052 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.|
|WB||1/2000. Detects a band of approximately 11 kDa.|
|ChIP||Use at an assay dependent concentration. PubMed: 20829797|
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
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
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ab9052 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 ab9052 at 0.5µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse 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.
Histone H4 (di methyl K20) was immunoprecipitated using 0.5mg HeLa whole cell extract, 5µg of Rabbit polyclonal to Histone H4 (di methyl K20) 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, Hela whole cell extract 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 ab9052.
Secondary: Clean blot (HRP conjugate) at 1/1000 dilution.
Band: 14kDa: Histone H4 (di methyl K20).
All lanes : Anti-Histone H4 (di methyl K20) antibody - ChIP Grade (ab9052) at 1/2000 dilution
Lane 1 : yH4 17-24 peptide
Lane 2 : Unmodified yH4 17-24 peptide
Lane 3 : Mono methyl K20 yH4 17-24 peptide
Lane 4 : Di methyl K20 yH4 17-24 peptide
Lane 5 : Tri methyl K20 yH4 17-24 peptide
Blocking peptides at 1 µg/ml per lane.
Performed under reducing conditions.
All lanes : Upper blot - Histone H4 antibody (gift of A. Verreault)
Lower blot - Rabbit polyclonal to Histone H4 di methyl K20 (ab9052) at 1/2000
Lane 1 : S.cerevisiae extract
Lane 2 : S.pombe extract
Lane 3 : S.pombe rH4
Lane 4 : Drosophila rH4
Lane 5 : Histones (Roche)
Di methylation at K20 is seen in S. pombe and in calf thymus (Roche). S. cerevisiae lacks K20 (di) methylation.
IHC image of ab9052 staining in human pancreas adenocarcinoma 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 ab9052, 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.
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.
This product has been referenced in:
- Alyodawi K et al. Compression of morbidity in a progeroid mouse model through the attenuation of myostatin/activin signalling. J Cachexia Sarcopenia Muscle 10:662-686 (2019). Read more (PubMed: 30916493) »
- Wen HJ et al. Myeloid Cell-Derived HB-EGF Drives Tissue Recovery After Pancreatitis. Cell Mol Gastroenterol Hepatol 8:173-192 (2019). Read more (PubMed: 31125624) »