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AbFlex® Histone H3K9ac antibody (rAb)
| RRID: AB_2793775 |
| 商品コード: 91103 | Format: 100 µg | ¥72,000 | Buy |
| 商品コード: 91104 | Format: 10 µg | ¥20,000 | Buy |
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Applications
Application Notes
Applications Validated by Active Motif:
ChIP-Seq: 4 µg each
ICC/IF: 1 - 2 µg/ml
WB*: 0.2 - 2 µg/ml
Bead-based ELISA: 1.5 - 40 µg/ml
CUT&Tag: 1 µg 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.
AbFlex® recombinant antibodies are genetically derived from DNA sequences of parental hybridoma clones. For details on the parental antibody clone, see Catalog No. 61251.
Immunogen
This antibody was raised against a peptide containing acetyl-lysine 9 of human Histone H3.
Buffer
Purified IgG in 140 mM Hepes, pH 7.5, 70 mM NaCl, 32 mM NaOAc, 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.
AbFlex® Histone H3K9ac antibody (rAb) tested by ChIP-Seq.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 30 µg of chromatin from PC9 cell line and 4 µg of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 22 million sequence tags were mapped to identify Histone H3K9ac binding sites. The image shows binding across a region of chromosome 1.
AbFlex® Histone H3K9ac antibody (rAb) tested by immunofluorescence.
HeLa cell stained with 2µg /mL of AbFlex® Histone H3K9ac antibody (rAb) followed by anti-mouse-IgG-488.
AbFlex® Histone H3K9ac antibody (rAb) tested by Western blot.
HeLa nuclear extract (20 µg per lane) probed with AbFlex® Histone H3K9ac antibody (0.5 µg/ml dilution). Lane 1: untreated cells. Lane 2: cells treated with sodium butyrate.
AbFlex® Histone H3K9ac antibody (rAb) tested by Luminex bead-based specificity analysis.
Luminex bead-based specificity analysis was used to confirm the specificity of AbFlex® Histone H3K9ac antibody (rAb) antibody for acetyl-lysine 9 H3. Peptides corresponding to regions around major sites of histone H3 acetylation or other acetyl-lysine peptides were conjugated to MagPlex Luminex beads and incubated with various amounts of AbFlex®Histone H3K9ac antibody (rAb). Peptide-bound antibody was detected with anti-mouse IgG-Phycoerythrin and read in a Luminex instrument.
Luminex® is a registered trademark of Luminex Corporation.
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 H3K9ac 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.
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.Acetylation of histone H3 Lys9 is associated with transcriptional activation of the genes.
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 H3K9ac 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
