Histone H3K9ac antibody (pAb)

RRID: AB_2561017
Catalog No: 39137 Format: 100 µl $455 Buy
Catalog No: 39038 Format: 50 µl $265 Buy
Catalog No: 39138 Format: 10 µl $110 Buy

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

Applications

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

Application Notes


Validated Applications:
ChIP: 10 µl per ChIP
ChIP-Seq: 5 µl each
WB: 1:1,000 - 1:2,000 dilution*

Published Applications:
ChIP / ChIP-Seq
WB
IF
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. This Histone H3 acetyl Lys9 antibody was validated for ChIP-Seq in the study (see references).

*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.

For Histone H3K9ac, we also offer AbFlex® Histone H3K9ac Recombinant Antibody (rAb).  For details, see Catalog No. 91103.

Immunogen

This Histone H3 acetyl Lys9 antibody was raised against a peptide including acetyl-lysine 9 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. 39917) of this antibody that was purified by Protein A Chromatography is also available.

References

 

Histone H3 acetyl Lys9 antibody tested by ChIP analysis.
Chromatin IP performed using the ChIP-IT® Express Kit (Catalog No. 53008) and HeLa Chromatin (1.5 x 106 cell equivalents per ChIP) using 10 µl of Histone H3 acetyl Lys9 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.

Histone H3 acetyl Lys9 antibody tested by Western blot.
HeLa acid extract probed with Histone H3 acetyl Lys9 polyclonal antibody (1:2,000 dilution).
    Lane 1: No treatment.
    Lane 2: Cells treated with sodium butyrate.

Histone H3 acetyl Lys9 antibody specificity tested by peptide array analysis.
Peptide array analysis was used to confirm the specificity of this antibody for its intended modification. Histone H3 acetyl Lys9 antibody was applied at a dilution of 1:3,000 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.

Histone H3 acetyl Lys9 antibody tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 acetyl Lys9 antibody for acetyl Lys9 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). 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 Lys9 can also be mono-, di- or trimethylated. The methylation of this residue is often associated with transcriptional repression. However, acetylation of histone H3 Lys9 is associated with transcriptional activation of the genes.

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 Lys9 antibody.

qPCR Primers

Active Motif's Human Positive Control Primer Set ACTB-2 (Cat. No. 71005) and Human Negative Control Primer Set 1 (Cat. No. 71001) have been validated for performing qPCR and endpoint PCR when this antibody has been used for ChIP on human samples. For ChIP experiments with this antibody on mouse samples, the Mouse Positive Control Primer Set Actb-2 (Cat. No. 71017) and Mouse Negative Control Primer Set 1 (Cat. No. 71011) have been validated for both qPCR and endpoint PCR.

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 H3K9ac antibody (pAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 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;
  • WB = Western Blotting