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AbFlex® Histone H3K27ac antibody (rAb)
| RRID: AB_2793797 |
| 商品コード: 91193 | Format: 100 µg | ¥72,000 | Buy |
| 商品コード: 91194 | Format: 10 µg | ¥20,000 | Buy |
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Applications
Application Notes
Applications Validated by Active Motif:
ChIP-Seq: 4 µg per ChIP
WB*: 0.2 - 2 µg/ml
Bead-based ELISA: 1 - 10 µg/ml
CUT&Tag: 1 µl per 50 µl reaction
CUT&RUN: 1 µl per 50 µl reaction
*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.
We offer a polyclonal version of Histone H3K27ac. For details, see Catalog No. 39685.
Published Applications
The following applications have been published using this antibody. Unless noted above, Active Motif may not have validated the antibody for use in these applications:
ChIP-qPCR
View publications that use Active Motif products & services here. Enter the product number(s) to see publications & applications that use this antibody.
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, 32 mM NaOAc, 0.035% sodium azide, 30% glycerol. Sodium azide is highly toxic.
AbFlex® Histone H3K27ac antibody 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.
AbFlex® Histone H3K27ac antibody 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 specificity.
Luminex bead-based analysis confirms specificity of AbFlex Histone H3K27ac antibody for H3K27ac. Peptides corresponding to regions around major sites of histone H3 acetylation or other acetyl-lysine peptides were conjugated to MagPlex Luminex beads, incubated with a range of AbFlex Histone H3K27ac antibody, and peptide-bound antibody was detected with anti-mouse IgG-PE (read in a Luminex instrument).
AbFlex® Histone H3K27ac antibody tested by CUT&Tag
CUT&Tag was performed using 250,000 K562 cells and sequenced using 38 base-pair, paired-end reads on the Illumina NextSeq 500/550. Data was collected from 31 million reads, and Histone H3K27ac data is shown for Chromosome 1.
AbFlex® H3K27ac antibody (rAb) tested by CUT&RUN
CUT&RUN was performed using 500,000 K562 nuclei (TOP) or 500,000 K562 cells (BOTTOM) and sequenced using 38 base-pair, paired-end reads on the Illumina NovaSeq. Data was collected from 24 million reads (TOP) and 26 million reads (BOTTOM). H3K4me3 data is shown for Chromosome 19.
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, a Biotinylation Tag 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).
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;
- ChIP = ChIP Sequencing;
- CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
- CUT&Tag = Cleavage Under Targets and Tagmentation;
- 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;
- NEU = Neutralizing;
- WB = Western Blotting
