AbFlex Histone H3K9me0 antibody (rAb)

Host / Isotype

Mouse / IgG2a

Reactivity

Human, Wide Range Predicted

Applications

ChIP, ChIP-Seq, ELISA, ICC, IF, WB

Cat No : 91155,91156

Synonyms

Histone H3, histone methylation, transcriptional activation chromatin, antibody, antibodies, monoclonal, H3K9me0, lysine 9, methyl, mAb, western blotting, wb, sample, recombinant, rAb, rec Ab, AbFlex

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Validation Data Gallery

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  AbFlex Histone H3K9me0 Antibody (rAb) tested by ChIP-Seq. ChIP was performed using the ChIP-IT High Sensitivity Kit (Cat. No. 53040) with 30 ug of chromatin from PC9, lung adenocarcinoma, cells and 4 ug of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 30 million sequence tags were mapped to identify Histone H3K9me0 binding sites. The image shows binding across a region of chromosome 17.

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  AbFlex Histone H3K9me0 Antibody (rAb) tested by immunofluorescence. HeLa cell stained with 2ug /mL of AbFlex Histone H3K9me0 Antibody (rAb followed by anti-mouse-IgG-488.

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  AbFlex Histone H3K9me0 antibody (rAb) tested by Western blot. HeLa nuclear extract (20 ug per lane) probed with AbFlexTM Histone H3K9me0 antibody (0.2 ug/ml dilution).

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Product Information

• Tested Applications: ChIP, ChIP-Seq, ELISA, ICC, IF, WB

  Applications Validated by Active Motif: ICC/IF: 1 - 2 ug/ml WB*: 0.1 - 1 ug/ml Bead-based ELISA: 0.5 - 10 ug/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.

• Tested Reactivity: Human, Wide Range Predicted
• Host / Isotype: Mouse / IgG2a
• Class: Recombinant
• Type: Antibody
• Modification: Methylated
• Immunogen: This antibody was raised against a synthetic branched peptide corresponding to amino acids surrounding Lys9 of human Histone H3.
• Full Name: AbFlex Histone H3K9me0 antibody (rAb)
• Synonyms: Histone H3, histone methylation, transcriptional activation chromatin, antibody, antibodies, monoclonal, H3K9me0, lysine 9, methyl, mAb, western blotting, wb, sample, recombinant, rAb, rec Ab, AbFlex
• Molecular weight: 17 kDa
• GenBank accession number: NP_003522
• RRID: AB_2793790
• Purification Method: Ni-NTA
• Buffer: Purified IgG in 200 mM Hepes pH 7.5, 100 mm NaCl, 50 mM NaOAc, 30% glycerol and 0.035% sodium azide. 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 H3K9me0 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 basic building block of chromatin. Histones are 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. Methylation of lysines occur as four states: unmethylated (me0), monomethyl (me1), dimethyl (me2) and trimethyl (me3). The me0 state of lysine is recognized as biologically relevant and a number of proteins containing PhD fingers, ADD and WD40 domains are known to associate with unmodified lysines.