AbFlex® Histone H3K27ac antibody (rAb)

RRID: AB_2793797
Catalog No: 91193 Format: 100 µg ¥62,000 Buy
Catalog No: 91194 Format: 10 µg ¥17,000 Buy

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Antibody Type:
Recombinant
Isotype:
IgG2a
Purification:
Protein A Chromatography
Host:
Mouse
Molecular Weight:
17 kDa
Reactivity:
Human, Wide Range Predicted

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Enzyme-linked Immunosorbent Assay Validated

Application Notes


Validated Applications:
ChIP-Seq: 4 µg per ChIP
WB*: 0.2 - 2 µg/ml
Bead-based ELISA: 1 - 10 µg/ml

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

Buffer

Purified IgG in 140 mM Hepes, pH 7.5, 70 mM NaCl, 35 mM NaOAc, 0.035% sodium azide, 30% glycerol.

View our Guide to Histone Modifications and Biological Function.

 

AbFlex® Histone H3K27ac (rAb) tested by ChIP-Seq.
Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 30 µg of chromatin from HAP1 cells and 4 µg Histone H3K27ac antibody. ChIP DNA was sequenced on the Illumina HiSeq and 17.8 million sequence tags were mapped to identify H3K27ac binding sites. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

AbFlex® Histone H3K27ac antibody (rAb) tested by Western Blot
HeLa cells were treated with 5 mM sodium butyrate for 19 hrs. (Lane 2) or untreated (Lane 1). Following treatment, 20 µg of HeLa cell nuclear extract was loaded on SDS-PAGE gel and probed with 1 µg/ml AbFlex Histone H3K27ac antibody. MW: 17 kDa

AbFlex Histone H3K27ac antibody, tested by Dot Blot
Dot blot analysis was used to confirm the specificity of AbFlex Histone H3K27ac antibody for acetylated H3K27. Recombinant Histone Proteins were spotted onto PVDF and probed with the antibody at 1 µg/ml. The amount of protein (picomoles) spotted is indicated next to each row. Columns as follows: 1) H3K37ac 2) Histone H3K36ac 3) Histone H3K9ac 4) Histone H3K14ac 5) Histone H3K18ac 6) Histone H3K23ac 7) Histone Unmod H3K27 8) Histone H3K27ac 9) Histone H4K5ac 10) Histone H4K8ac 11) Histone H4K12ac 12) Histone H4K16ac

Background

AbFlex® antibodies are recombinant antibodies (rAbs) that have been generated using defined DNA sequences to produce highly specific, reproducible antibodies. Each AbFlex antibody contains a 6xHis-Tag, an avidin tag sequence for enzymatic biotin conjugation using the biotin ligase, BirA, and a sortase recognition motif (LPXTG) to attach a variety of labels directly to the antibody including fluorophores, enzymatic substrates (HRP, AP), peptides, drugs as well as solid supports.

AbFlex® Histone H3K27ac antibody was expressed as full-length IgG with mouse immunoglobulin heavy and light chains (IgG2a isotype) in mammalian 293 cells.
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.

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 AbFlex® Histone H3K27ac antibody (rAb).

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 AbFlex® Histone H3K27ac antibody (rAb).

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;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting