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Histone H4 acetyl Lys16 antibody (pAb)
| Catalog No: 39167 | Format: 200 µl | $340 | Buy Now |
| Catalog No: 39168 | Format: 10 µl | $70 | Buy Now |
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Applications Table
| Western Blot | Chromatin IP | ChIP-Seq / ChIP-chip |
IF |
|---|---|---|---|
| 1:1,000 - 1:5,000 | 3 - 10 µl per ChIP | modENCODE validated. | Not recommended. |
for Histone H4 acetyl Lys16 antibody (pAb)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). 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; these modifications play a major role in regulating gene expression.
Lysine N-ε-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. NoRC is a SNF2h-containing chromatin remodeling complex. The bromodomain of TIP5, the large subunit of NoRC, interacts with acetylated Histone H4 Lys16, (H4AcK16) and cooperates with an adjacent PHD finger to recruit histone deacetylases and DNA methyltransferases to rDNA, leading to the silencing of rDNA.
for Histone H4 acetyl Lys16 antibody (pAb)Immunogen
This Histone H4 acetyl Lys16 antibody was raised against a peptide including acetyl-lysine 16 of histone H4.
for Histone H4 acetyl Lys16 antibody (pAb)Application Notes
This Histone H4 acetyl Lys16 antibody (H4K16ac) has been validated for use in chromatin IP (3 - 10 µl per ChIP), ChIP-Seq & ChIP-chip (5 µl each) and Western blotting (1:1,000 - 1:5,000 dilution). Specificity for acetyl-lysine 16 was confirmed by dot blot and peptide inhibition with non-modified histone H4 peptide or peptides acetylated at lysines 5, 8 or 12. Because this modification is conserved from yeast to human, a wide range of reactivity is expected. Overnight incubation with this antibody is recommended for Western blotting. Due to cytoplasmic staining, this product is not recommended for IF.
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 Lys16 antibody was validated for ChIP-Seq in the study (see References).
for Histone H4 acetyl Lys16 antibody (pAb)Buffer
Rabbit serum containing 30% glycerol and 0.035% sodium azide.
for Histone H4 acetyl Lys16 antibody (pAb)Positive Control
Active Motif's HeLa acid extract (Sodium butyrate treated) (Catalog No. 36202) can be used as a positive control for Histone H4 acetyl Lys16 antibody.
Chromatin IP Assays
Active Motif has a variety of Chromatin Immunoprecipitation Kits for every need.
Please visit our Chromatin IP Page to learn about ChIP products designed for use with Histone H4 acetyl Lys16 antibody (pAb).
for Histone H4 acetyl Lys16 antibody (pAb)References
This antibody has been used as described below:
Histone H4 acetyl Lys16 antibody (pAb) tested by Western blot.
HeLa acid extract probed with Histone H4 acetyl Lys16 polyclonal antibody (1:1,000 dilution).
Lane 1: No treatment.
Lane 2: Cells treated with sodium butyrate.
Histone H4 acetyl Lys16 antibody (pAb) tested by ChIP.
ChIP performed on budding yeast chromatin.
Lane 1: ChIP performed using Histone H4 acetyl Lys16 pAb (3 µl).
Lane 2: Input DNA PCR control.
Multiplex PCR was performed using primers specific for the yeast SCC2 gene (SCC2) and an internal PCR control using primers specific for a sub-telomeric sequence (TEL).
Histone H4 acetyl Lys16 antibody (pAb) tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H4 acetyl Lys16 pAb for histone H4 acetyl Lys16. Acetylated peptides corresponding to the immunogen and related H4 sequences were spotted onto PVDF and probed with the antibody at 1:5,000. The amount of peptide (picomoles) spotted is indicated next to each row.
Lane 1: Acetyl-Lys5 peptide.
Lane 2: Unmodified Lys5 peptide.
Lane 3: Acetyl-Lys8 peptide.
Lane 4: Unmodified Lys8 peptide.
Lane 5: Acetyl-Lys12 peptide.
Lane 6: Unmodified Lys12 peptide.
Lane 7: Acetyl-Lys16 peptide.
Lane 8: Unmodified Lys16 peptide.
Histone H4 acetyl Lys16 antibody (pAb) tested by ChIP analysis.
Chromatin IP performed using the ChIP-IT® Express Kit (Catalog No. 53008) and HeLa Chromatin (1.5 x 10^6 cell equivalents per ChIP) using 10 µl of Histone H4 acetyl Lys16 pAb 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.
Histone H4 acetyl Lys16 antibody specificity tested by peptide array analysis.
Peptide array analysis was used to confirm the specificity of this antibody for its intended modification. Histone H4 acetyl Lys16 antibody was applied at a dilution of 1:10,000 to Active Motif's MODified™ Histone Peptide Array (Catalog No. 13001). The arrays were scanned with ArrayAnalysis Software 7 and the results plotted. Specificity data is shown for the most reactive peptides and those related to the immunogen. In the array, the acetyl Lys16 modification is found on peptides with two different backbone sequences (H4 amino acids 1-19 and amino acids 11-30) so reactive and related peptides for both peptides is shown.
Array Data File
for Histone H4 acetyl Lys16 antibody (pAb)Storage
Antibodies in solution can be stored at -20°C for 2 years. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
for Histone H4 acetyl Lys16 antibody (pAb)Guarantee
This product is guaranteed for 6 months from date of receipt.
This product is for research use only and is not for use in diagnostic procedures.
Application Key
- ChIP = Chromatin Immunoprecipitation;
- DB = Dot Blot;
- ELISA = Enzyme-linked Immunosorbent Assay;
- EMSA = Electrophoretic Mobility Shift Assay
- FACS = Flow Cytometry;
- ICC = Immunocytochemistry;
- IF = Immunofluorescence;
- IHC = Immunohistochemistry;
- IP = Immunoprecipitation;
- MeDIP = Methyl-DNA Immunoprecipitation;
- WB = Western Blotting
