Chromatin Immunoprecipitation (ChIP)
(Histone H4 acetyl Lys12 antibody tested by ChIP analysis. Chromatin IP performed using the ChIP-IT Express Kit and HeLa Chromatin (1.5 x 106 cell equivalents per ChIP) using 10 ul of Histone H4 acetyl Lys12 antibody or the equivalent amount of rabbit IgG as a negative control. Real time, quantitative PCR (RT-qPCR) was performed on DNA purified from each of the ChIP reactions using a primer pair specific for the indicated gene. Data are presented as Fold Enrichment of the ChIP antibody signal versus the negative control IgG using the ddCT method.)
Rabbit Histone H4K12ac Polyclonal Antibody | anti-H4K12ac antibody
Histone H4K12ac antibody (pAb)
ChIP: 2-8ul per ChIP
ChIP-Seq: 5ul each
ICC/IF: 1:500-1:2,000 dilution
WB: 1:5,000-1:20,000 dilution
The modENCODE and NIH Roadmap Epigenomics Mapping Consortiums have implemented rigorous standardization criteria for all assays and reagents to be used. As part of this initiative, antibody specificity testing and the ability of the antibodies to work in ChIP-Seq were assessed in a large-scale study. This Histone H4 acetyl Lys12 antibody was validated for ChIP-Seq in the study (see reference).
Chromatin Immunoprecipitation (ChIP)
(Histone H4 acetyl Lys12 antibody tested by ChIP analysis. Chromatin IP performed using the ChIP-IT Express Kit and HeLa Chromatin (1.5 x 106 cell equivalents per ChIP) using 10 ul of Histone H4 acetyl Lys12 antibody or the equivalent amount of rabbit IgG as a negative control. Real time, quantitative PCR (RT-qPCR) was performed on DNA purified from each of the ChIP reactions using a primer pair specific for the indicated gene. Data are presented as Fold Enrichment of the ChIP antibody signal versus the negative control IgG using the ddCT method.)
Chromatin Immunoprecipitation (ChIP)
(Histone H4 acetyl Lys12 antibody tested by ChIP analysis. Chromatin IP performed using the ChIP-IT Express Kit and HeLa Chromatin (1.5 x 106 cell equivalents per ChIP) using 10 ul of Histone H4 acetyl Lys12 antibody or the equivalent amount of rabbit IgG as a negative control. Real time, quantitative PCR (RT-qPCR) was performed on DNA purified from each of the ChIP reactions using a primer pair specific for the indicated gene. Data are presented as Fold Enrichment of the ChIP antibody signal versus the negative control IgG using the ddCT method.)
Immunofluorescence (IF)
(Histone H4 acetyl Lys12 antibody tested by immunofluorescence. Staining of HeLa cells with Histone H4 acetyl Lys12 antibody (1:500 dilution, top panel) and DAPI (bottom panel).)
Immunofluorescence (IF)
(Histone H4 acetyl Lys12 antibody tested by immunofluorescence. Staining of HeLa cells with Histone H4 acetyl Lys12 antibody (1:500 dilution, top panel) and DAPI (bottom panel).)
Western Blot (WB)
(Histone H4 acetyl Lys12 antibody tested by Western blot. HeLa acid extract probed with Histone H4 acetyl Lys12 polyclonal antibody (1:10,000 dilution). Lane 1: No treatment. Lane 2: Cells treated with sodium butyrate.)
Western Blot (WB)
(Histone H4 acetyl Lys12 antibody tested by Western blot. HeLa acid extract probed with Histone H4 acetyl Lys12 polyclonal antibody (1:10,000 dilution). Lane 1: No treatment. Lane 2: Cells treated with sodium butyrate.)
Dot Blot (DB)
(Specificity Data. Dot blot was used to confirm the specificity of Histone H4K12ac antibody for H4 acetyl Lys12. Acetylated peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed at a dilution of 1:5,000. The amount of peptide (picomoles) spotted is indicated next to each row. Column 1: unmod H4K8. Column 2: H4K5ac. Column 3: H4K8ac. Column 4: H4K12ac. Column 5: unmod H4K16. Column 6: H4K16ac. Column 7: H4K20ac. Column 8: unmod H4K31. Column 9: HrK31ac. Column 10: H4K59ac. Column 11: H4K91ac.)
Dot Blot (DB)
(Specificity Data. Dot blot was used to confirm the specificity of Histone H4K12ac antibody for H4 acetyl Lys12. Acetylated peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed at a dilution of 1:5,000. The amount of peptide (picomoles) spotted is indicated next to each row. Column 1: unmod H4K8. Column 2: H4K5ac. Column 3: H4K8ac. Column 4: H4K12ac. Column 5: unmod H4K16. Column 6: H4K16ac. Column 7: H4K20ac. Column 8: unmod H4K31. Column 9: HrK31ac. Column 10: H4K59ac. Column 11: H4K91ac.)
Background: Histone H4 is one of the core components of the nucleosome. The nucleosome is the smallest subunit of chromatin and consists of 147 base pairs of DNA wrapped around an octamer of core histone proteins (two each of Histone H2A, Histone H2B, Histone H3 and Histone H4). Histone H1 is a linker histone, present at the interface between the nucleosome core and DNA entry/exit points; it is responsible for establishing higher-order chromatin structure. Chromatin is subject to a variety of chemical modifications, including post-translational modifications of the histone proteins and the methylation of cytosine residues in the DNA. Reported histone modifications include acetylation, methylation, phosphorylation, ubiquitylation, glycosylation, ADP-ribosylation, carbonylation and SUMOylation; they play a major role in regulating gene expression. Lysine N-epsilon-acetylation is a dynamic, reversible and tightly regulated protein and histone modification that plays a major role in chromatin remodeling and in the regulation of gene expression in various cellular functions. The acetylation of histone H4 Lys12 is carried out by several histone acetyltransferases (HATs). Histone H4 Lys12 acetylation by TIP60 occurs in the nucleus, whereas Hat1-mediated acetylation of histone H4 Lys12 occurs in the cytoplasm, indicating the spatial regulation of histone acetylation. The acetylation of histone H4 Lys12 residue is commonly associated with transcriptional activation.
NCBI and Uniprot Product Information
Uniprot Description
Function: Core 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.
Subunit structure: The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA.
Subcellular location: Nucleus. Chromosome.
Post-translational modification: Acetylation 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. Ref.19 Ref.20 Ref.21 Ref.23 Ref.24 Ref.28Phosphorylated by PAK2 at Ser-48 (H4S47ph). This phosphorylation increases the association of H3.3-H4 with the histone chaperone HIRA, thus promoting nucleosome assembly of H3.3-H4 and inhibiting nucleosome assembly of H3.1-H4. Ref.28 Ref.36Ubiquitinated 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). Ref.27 Ref.30Sumoylated, which is associated with transcriptional repression. Ref.22Crotonylation (Kcr) is specifically present in male germ cells and marks testis-specific genes in post-meiotic cells, including X-linked genes that escape sex chromosome inactivation in haploid cells. Crotonylation marks active promoters and enhancers and confers resistance to transcriptional repressors. It is also associated with post-meiotically activated genes on autosomes. Ref.35
Sequence similarities: Belongs to the histone H4 family.
Sequence caution: The sequence AAI28106.1 differs from that shown. Reason: Frameshift at position 3.
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Product Notes
The H4K12ac hist1h4a (Catalog #AAA388352) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Histone H4K12ac antibody (pAb) reacts with Budding Yeast, Human, Mouse, Wide Range Predicted and may cross-react with other species as described in the data sheet. AAA Biotech's Histone H4K12ac can be used in a range of immunoassay formats including, but not limited to, Chromatin Immunoprecipitation (ChIP), Chromatin Immunoprecipitation-Seq (ChIP-Seq), Dot Blot (DB), Immunocytochemistry (ICC), Immunofluorescence (IF), Western Blot (WB). Applications Validated: ChIP: 2-8ul per ChIP ChIP-Seq: 5ul each ICC/IF: 1:500-1:2,000 dilution WB: 1:5,000-1:20,000 dilution The modENCODE and NIH Roadmap Epigenomics Mapping Consortiums have implemented rigorous standardization criteria for all assays and reagents to be used. As part of this initiative, antibody specificity testing and the ability of the antibodies to work in ChIP-Seq were assessed in a large-scale study. This Histone H4 acetyl Lys12 antibody was validated for ChIP-Seq in the study (see reference). Researchers should empirically determine the suitability of the H4K12ac hist1h4a for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Histone H4K12ac, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
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