Histone H3K9ac antibody (mAb)

Clone: 1B10
Catalog No: 61251 Format: 100 µg $445 Buy
Catalog No: 61952 Format: 50 µg $260 Buy
Catalog No: 61252 Format: 10 µg $105 Buy

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Antibody Type:
Monoclonal
Isotype:
IgG3
Purification:
Protein A Chromatography
Host:
Mouse
Molecular Weight:
17
Reactivity:
Human, Mouse, Other (Wide Range)

Applications

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

Application Notes



Validated Applications:
ChIP: 2 - 5 µg per ChIP
ChIP-Seq: 5 µg each
WB: 0.2 - 2 µg/ml dilution

Published Applications:
ChIP
WB
See references for more information. Individual optimization may be required. NGS-QC® certification. This antibody has been processed by the NGS-QC® generator. For additional details, click here.

Immunogen

The Histone H3 acetyl Lys9 antibody was raised against a peptide including acetyl-lysine 9 of human Histone H3.

Buffer

Purified IgG in 0.1M Tris, pH 8.0, 1 M NaCl with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

 
Histone H3K9ac antibody (mAb) tested by ChIP-Seq.

Histone H3 acetyl Lys9 antibody (mAb) tested by ChIP-Seq.
ChIP was performed using the ChIP-IT High Sensitivity® Kit (Cat. No. 53040) with chromatin from rat brain FFPE sections and 5 µl of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 5.2 million unique sequence tags were mapped to identify H3K9ac binding sites. The image on the left shows binding across the entire length of chromosome 20 and the data indicates good correlation of gene density and H3K9ac signal. The image on the right is zoomed in to show H3K9ac at the promoter of two genes.

Histone H3K9ac antibody (mAb) tested by ChIP.

ChIP of Histone H3 acetyl Lys9 mAb.
Chromatin IP performed using the ChIP-IT® Express Kit (Cat. No. 53008) and HeLa chromatin (1.5 x 106 cell equivalents per ChIP) using 3 µg of Histone H3 acetyl Lys9 antibody or the equivalent amount of mouse 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. Data are presented as Fold Enrichment of the ChIP antibody signal versus the negative control IgG using the ddCT method.

Histone H3K9ac antibody (mAb) tested by immunofluorescence.

Histone H3 acetyl Lys9 antibody tested by immunofluorescence.
Detection of Histone H3 acetyl Lys9 by immunofluorescence. HeLa cells were stained with Histone H3 acetyl Lys9 antibody at a dilution of 2 µg/ml. Top panel: DAPI. Middle panel: Histone H3 acetyl Lys9 antibody staining. Bottom panel: merge.

Histone H3K9ac antibody (mAb) tested by Western blot.

Histone H3 acetyl Lys9 antibody (mAb) tested by Western blot.
HeLa nuclear extract (20 µg per lane) probed with Histone H3 acetyl Lys9 antibody (2 µg per ml).
Lane 1: no treatment. Lane 2: cells treated with sodium butyrate.

Histone H3K9ac antibody (mAb) tested by dot blot analysis.

Histone H3 acetyl Lys9 mAb tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 acetyl Lys9 mAb for acetyl Lys9 of histone H3. Acetylated peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed with the antibody at a dilution of 2 µg per ml. The amount of peptide (picomoles) spotted is indicated next to each row. Top 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. Bottom row: Lane 1: acetyl-Lys56 peptide. Lane 2: acetyl-Lys64 peptide. Lane 3: H4 acetyl-Lys91 peptide. Lane 4: acetyl Lys37 peptide. Lane 5: acetyl-Lys36 peptide. Lane 6: acetyl-Lys4 peptide. Lane 7: acetyl-Lys122 peptide. Lane 8: acetyl-Lys79 peptide.

Background

Histone H3 is one of the core components of the nucleosome, the basic building block of chromatin. Histones are 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. Acetylation of Lys9 is associated with trascriptional activation of genes.

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 (mAb).

Bridging Antibody to Improve ChIP, MeDIP and IP

Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use Histone H3K9ac antibody (mAb).

Compatible Secondary Antibodies

Active Motif has developed a variety of high-quality secondary antibodies for most applications, with anti-rabbit and anti-mouse conjugates to a broad line of high-quality fluorescent molecules, as well as horse radish peroxidase (HRP). Visit our Secondary Antibody Conjugates page to find the perfect secondaries for immunofluorescence experiments with Histone H3K9ac antibody (mAb).

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