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Antibodies

Histone H3K27ac antibody (mAb)

Clone: MABI 0309
Catalog No: 39685 Format: 100 µg $420 Buy Now

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Antibody Type:Monoclonal
Isotype:IgG1
Purification:Protein G Chromatography
Host:Mouse
Molecular Weight:17 kDa
Reactivity:Human, Other (Wide Range)

Applications

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

for Histone H3 acetyl Lys27 antibody (mAb) (Clone MABI 0309)Application Notes


Validated Applications:
ChIP: 5 - 10 µg per ChIP
ChIP-Seq: 5 - 10 µg each
ICC/IF: 1 - 2 µg/ml dilution
WB: 0.5 - 2 µl/ml dilution

ChIP-Seq validation was performed by Active Motif's Epigenetics Services and the complete data set is available in the UCSC Genome Browser by clicking here.

NGS-QC® certification. This antibody has been processed by the NGS-QC® generator. For additional details, click here..

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

for Histone H3 acetyl Lys27 antibody (mAb) (Clone MABI 0309)Immunogen

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

for Histone H3 acetyl Lys27 antibody (mAb) (Clone MABI 0309)Buffer

PBS pH 7.5 containing 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

for Histone H3 acetyl Lys27 antibody (mAb) (Clone MABI 0309)References

 
Histone H3K27ac antibody (mAb) tested by ChIP-Seq.

Histone H3K27ac antibody (mAb) tested by ChIP-Seq.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 15 ug of chromatin from a human medulloblastoma cell line and 4 µg of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 17 million sequence tags were mapped to identify Histone H3K27ac binding sites. The image shows binding across a region of chromosome 7. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

Histone H3K27ac antibody (mAb) tested by immunofluorescence.

Histone H3 acetyl Lys27 mAb tested by immunofluorescence.
Staining of HeLa cells with Histone H3 acetyl Lys27 mAb (1 µg/ml, top panel) and DAPI (middle panel), and a merge of both images (bottom panel).

Histone H3K27ac antibody (mAb) tested by Western blot.

Histone H3 acetyl Lys27 mAb tested by Western blot.
HeLa nuclear extract (20 µg per lane) probed with Histone H3 acetyl Lys27 mAb (2 µg/ml dilution).
Lane 1: No treatment.
Lane 2: cells treated with sodium butyrate.

Histone H3K27ac antibody (mAb) tested by dot blot analysis.

Histone H3 acetyl Lys27 mAb tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 acetyl Lys27 mAb for acetyl-Lys27 histone H3. Acetylated peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed with the antibody at 0.2 µg/ml. The amount of peptide (picomoles) spotted is indicated next to each row.
Lane 1: acetyl-Lys4 peptide. Lane 2: acetyl-Lys9 peptide. Lane 3: acetyl-Lys14 peptide. Lane 4: acetyl-Lys18 peptide. Lane 5: acetyl-Lys23 peptide. Lane 6: acetyl-Lys27 peptide. Lane 7: acetyl-Lys36 peptide. Lane 8: acetyl-Lys37 peptide. Lane 9: acetyl-Lys64 peptide. Lane 10: acetyl-Lys79 peptide.

for Histone H3 acetyl Lys27 antibody (mAb) (Clone MABI 0309)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 27 of histone H3 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.

for Histone H3 acetyl Lys27 antibody (mAb) (Clone MABI 0309)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 mAb (Clone MABI 0309).

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 (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 H3K27ac 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 H3K27ac antibody (mAb).

for Histone H3 acetyl Lys27 antibody (mAb) (Clone MABI 0309)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.

for Histone H3 acetyl Lys27 antibody (mAb) (Clone MABI 0309)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
 

Technical Data Sheet

Histone H3K27ac antibody (mAb)
 

Data Thumbnails

Histone H3K27ac antibody (mAb) tested by ChIP-Seq.

ChIP-Seq of Histone H3K27ac mAb.
(Click image to enlarge and see details.)

Histone H3K27ac antibody (mAb) tested by immunofluorescence.

Immunofluorescence stain of Histone H3K27ac mAb.
(Click image to enlarge and see details.)

Histone H3K27ac antibody (mAb) tested by Western blot.

Western blot of Histone H3K27ac mAb.
(Click image to enlarge and see details.)

Histone H3K27ac antibody (mAb) tested by dot blot analysis.

Dot blot of Histone H3K27ac mAb.
(Click image to enlarge and see details.)