Histone H3K27ac antibody (pAb)

RRID: AB_2614979
Catalog No: 39135 Format: 100 µl $445 Buy
Catalog No: 39136 Format: 10 µl $105 Buy

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Antibody Type:
Polyclonal
Isotype:
Serum
Purification:
None
Host:
Rabbit
Molecular Weight:
17 kDa
Reactivity:
Budding Yeast, Human, Wide Range Predicted

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Dot Blot Validated

Application Notes


Validated Applications:
ChIP: 3 µl per ChIP
ChIP-Seq: 5 µl each
WB: 1:1,000 - 1:5,000 dilution

Published Applications:
ChIP-Seq
ChIP
See references for more information. Individual optimization may be required.

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. Histone H3 acetyl Lys27 antibody was validated for ChIP-Seq in the study (see reference).
Note: many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western Blot.

Immunogen

This Histone H3 acetyl Lys27 antibody was raised against a peptide including acetyl-lysine 27 of histone H3.

Buffer

Rabbit serum containing 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, an IgG version (Catalog No. 39133) of this antibody that was purified by Protein A Chromatography is also available.

View our Guide to Histone Modifications and Biological Function.

References

 

Histone H3 acetyl Lys27 antibody tested by ChIP analysis.
Chromatin IP performed using the ChIP-IT® Express Kit (Catalog No. 53008) and HeLa Chromatin (2.1 x 106 cell equivalents per ChIP) using 3 µl of Histone H3 acetyl Lys27 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 primer pairs specific for the indicated gene as well as a negative control primer pair. Data are presented as Fold Enrichment of the ChIP antibody signal versus the negative control IgG using the ddCT method.

Histone H3 acetyl Lys27 antibody tested by Western blot.
HeLa acid extract probed with Histone H3 acetyl Lys27 polyclonal antibody (1:1,000 dilution).
    Lane 1: No treatment.
    Lane 2: Cells treated with sodium butyrate.

Histone H3 acetyl Lys27 pAb tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 acetyl Lys27 pAb for acetyl Lys27 histone H3. Acetylated peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed with the antibody at a dilution of 1:1,000. The amount of peptide (picomoles) spotted is indicated next to each row.
Lane 1: Acetyl-Lys9 peptide. Lane 2: Unmodified Lys9 peptide. Lane 3: Acetyl-Lys14 peptide. Lane 4: Unmodified Lys14 peptide. Lane 5: Acetyl-Lys18 peptide. Lane 6: Unmodified Lys18 peptide. Lane 7: Acetyl-Lys23 peptide. Lane 8: Unmodified Lys23 peptide. Lane 9: Acetyl-Lys27 peptide. Lane 10: Unmodified Lys27 peptide.

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). 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-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. Acetylation of histone H3 occurs at several different lysine positions in the histone tail, and is performed by Histone Acetyltransferases (HATs) such as CBP/p300. Acetylation of histone H3 at Lys27 is associated with transcriptional activation.

Histone H3K27 can also be mono-, di- or trimethylated by different histone methyltransferases, such as EZH2 or NSD3. While histone methylation can be associated with transcriptional activation or repression, methylation of Lysine 27 of histone H3 is mainly associated with transcriptional repression.

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 Lys27 antibody.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with Histone H3K27ac antibody (pAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

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
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting