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Histone H3 acetyl Lys56 antibody (pAb)
| Catalog No: 39281 | Format: 200 µl | $340 | Buy Now |
| Catalog No: 39282 | Format: 10 µl | $70 | Buy Now |
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Applications Table
| Western Blot | Chromatin IP |
|---|---|
| 1:500 - 1:2,500 | 2 - 5 µl per ChIP |
for Histone H3 acetyl Lys56 antibody (pAb)Background
Histone H3 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. Histone H3 Lys56 acetylation occurs normally during S phase, but disappears in G2. This modification persists in presence of DNA damage and also plays a role in nucleosome assembly. Rtt109 was shown to be the major histone acetyltransferase (HAT) for Lys56 acetylation.
for Histone H3 acetyl Lys56 antibody (pAb)Immunogen
This histone H3 acetyl Lys56 antibody was raised against peptide containing acetyl-Lys56 of yeast histone H3.
for Histone H3 acetyl Lys56 antibody (pAb)Application Notes
This Histone H3 acetyl Lys56 antibody (H3K56Ac) has been validated for use in Western blotting (1:500 - 1:2,500 dilution) and chromatin IP (5 - 10 µl per ChIP).
Detection of histone H3 by this antibody is completely abolished in extract from a yeast strain containing a substitution of arginine for lysine at position 56, as well as in extract from a strain deficient for the enzyme (Rtt109), which catalyzes acetylation of lysine 56.
for Histone H3 acetyl Lys56 antibody (pAb)Buffer
Purified rabbit IgG in 70 mM Tris (pH 8), 105 mM NaCl, 31 mM glycine, 0.07 mM EDTA, 30% glycerol and 0.035% sodium azide.
for Histone H3 acetyl Lys56 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 H3 acetyl Lys56 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 H3 acetyl Lys56 antibody (pAb).
Histone H3 acetyl Lys56 antibody tested by Western blot.
HeLa acid extract (10 µg/lane) was probed with Histone H3 acetyl Lys56 polyclonal antibody (1:3,500 dilution).
Lane 1: No treatment.
Lane 2: Cells treated with sodium butyrate.
Histone H3 acetyl Lys56 antibody tested by Western blot.
Acid extract of yeast cells probed with Histone H3 acetyl Lys56 pAb (1:1,400 dilution).
Histone H3 acetyl Lys56 pAb tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 acetyl Lys56 pAb for acetyl Lys56 histone H3. Acetylated peptides corresponding to the immunogen and related peptides 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-Lys56 peptide derived from budding yeast histone H3.
Lane 2: unmodified yeast histone H3 peptide.
Lane 3: acetyl-Lys56 peptide derived from human histone H3.
Lane 4: acetyl-Lys9 and acetyl-Lys14 peptide derived from human histone H3.
Lane 5: acetyl-Lys9 peptide derived from human histone H3.
Lane 6: unmodified peptide surrounding Lys9 derived from human histone H3.
Lane 7: acetyl-Lys14 peptide derived from human histone H3.
Lane 8: acetyl-Lys18 peptide derived from human histone H3.
Lane 9: acetyl-Lys23 peptide derived from human histone H3.
Lane 10: acetyl-Lys27 peptide derived from human histone H3.
Histone H3 acetyl Lys56 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 2 µl of Histone H3 acetyl Lys56 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.
for Histone H3 acetyl Lys56 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 H3 acetyl Lys56 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
