AbFlex Histone H3K27ac antibody (rAb)

Host / Isotype

Mouse / IgG2a

Reactivity

Human, Wide Range Predicted

Applications

ChIP-Seq, CUT&Tag, ELISA, WB

Cat No : 91193,91194

Synonyms

Histone H3, Histone acetylation, transcriptional activation, chromatin, nucleosome, antibody, antibodies, H3 lysine 27, chromatin immunoprecipitation, chip, recombinant antibody, recombinant, rAb, rec Ab,AbFlex

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Validation Data Gallery

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  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 ug of chromatin from HAP1 cells and 4 ug Histone H3K27ac antibody. ChIP DNA was sequenced on the Illumina HiSeq and 17.8 million sequence tags were mapped to identify H3K27ac binding sites.

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  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 ug of HeLa cell nuclear extract was loaded on SDS-PAGE gel and probed with 1 ug/ml AbFlex Histone H3K27ac antibody. MW: 17 kDa

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  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 ug/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

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  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.

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Product Information

• Tested Applications: ChIP-Seq, CUT&Tag, ELISA, WB

  Applications Validated by Active Motif: ChIP-Seq: 4 ug per ChIP WB*: 0.2 - 2 ug/ml Bead-based ELISA: 1 - 10 ug/ml CUT&Tag: 1 ul per 50 ul 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.

• Tested Reactivity: Human, Wide Range Predicted
• Host / Isotype: Mouse / IgG2a
• Class: Recombinant
• Type: Antibody
• Modification: Acetylated
• Immunogen: This antibody was raised against a synthetic peptide including acetyl-lysine 27 of human histone H3.
• Full Name: AbFlex Histone H3K27ac antibody (rAb)
• Synonyms: Histone H3, Histone acetylation, transcriptional activation, chromatin, nucleosome, antibody, antibodies, H3 lysine 27, chromatin immunoprecipitation, chip, recombinant antibody, recombinant, rAb, rec Ab,AbFlex
• Molecular weight: 17 kDa
• GenBank accession number: NP_003522
• RRID: AB_2793797
• Purification Method: Protein A Chromatography
• 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.
• 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.

Background Information

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.