Key features and details
- Rabbit polyclonal to Histone H4 (acetyl K12)
- Suitable for: ChIP-sequencing, WB, ICC/IF, Dot blot, ChIP
- Reacts with: Mouse, Human, Recombinant fragment
- Isotype: IgG
Product nameAnti-Histone H4 (acetyl K12) antibody
See all Histone H4 primary antibodies
DescriptionRabbit polyclonal to Histone H4 (acetyl K12)
Tested applicationsSuitable for: ChIP-sequencing, WB, ICC/IF, Dot blot, ChIPmore details
Species reactivityReacts with: Mouse, Human, Recombinant fragment
Synthetic peptide corresponding to Human Histone H4 (acetyl K12) conjugated to keyhole limpet haemocyanin.
Database link: P62805
- ChIP: HeLa cells. WB: HeLa whole cell and histone extracts. ICC/IF: HeLa cells.
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In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferPreservatives: 0.05% Sodium azide, 0.05% Proclin 300
Concentration information loading...
Our Abpromise guarantee covers the use of ab231678 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP-sequencing||Use at an assay dependent concentration.|
|ChIP||Use at an assay dependent concentration.|
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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All lanes : Anti-Histone H4 (acetyl K12) antibody (ab231678) at 1/500 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell extracts at 25 µg
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) histone extracts at 15 µg
Lane 3 : Recombinant H2A at 1 µg
Lane 4 : Recombinant H2B at 1 µg
Lane 5 : Recombinant H3 at 1 µg
Lane 6 : Recombinant H4 at 1 µg
Dilution buffer: TBS-Tween containing 5% skimmed milk.
ChIP assays were performed using HeLa (Human epithelial cell line from cervix adenocarcinoma) cells and optimized PCR primer sets for qPCR. ChIP was performed on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Image shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
ChIP was performed with 0.5 µg ab231678 on sheared chromatin from 1,000,000 HeLa (Human epithelial cell line from cervix adenocarcinoma) cells. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Image shows the peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (2C and D).
Dot Blot analysis was performed with peptides containing other histone modifcations and the unmodifed H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. ab231678 was used at a dilution of 1:1,000. Image shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac.
HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained for Histone H4 (acetyl K12) using ab231678 at a dilution of 1/200 in ICC/IF (Left panel).
Cells were fxed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. Secondary used is an Alexa Fluor®488-conjugated anti-Rabbit IgG. The right panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown at the bottom.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab231678 has not yet been referenced specifically in any publications.