AbFlex Histone H3K4me3 antibody (rAb)

Host / Isotype

Mouse / IgG2a

Reactivity

Human, Wide Range Predicted

Applications

ChIP, ChIP-Seq, DB, WB

Cat No : 91263,91264

Synonyms

Histone H3, histone methylation, transcriptional activation chromatin, antibody, antibodies, monoclonal, H3K4me3, AbFlex, recombinant antibodies

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Validation Data Gallery

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  AbFlex Histone H3K4me3 recombinant antibody (rAb) tested by ChIP-Seq Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT High Sensitivity Kit (Cat. No. 53040) with 30 ug of KMS11 cell chromatin and 4 ug of AbFlex Histone H3K4me3 antibody. ChIP DNA was sequenced on the Illumina NextSeq and 19.8 million sequence tags were mapped to identify Histone H3K4me3 binding sites on chromosome 7.

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  AbFlex Histone H3K4me3 recombinant antibody (rAb) tested by Western blot 20 ug of HeLa cell extract was run on SDS-PAGE and probed with AbFlex Histone H3K4me3 antibody at 2 ug/ml

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  AbFlex Histone H3K4me3 recombinant antibody (rAb) specificity tested by peptide array analysis. Peptide array analysis was used to confirm the specificity of this antibody for its intended modification. Histone H3K4me3 antibody was applied to Active Motif's MODified Histone Peptide Array (Catalog No. 13001). The arrays were scanned with ArrayAnalysis Software 7 and the results plotted. Specificity data is shown for the most reactive peptides and those related to the immunogen.

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Product Information

• Tested Applications: ChIP, ChIP-Seq, DB, WB

  Applications Validated by Active Motif: ChIP-Seq: 4 ug/ml WB: 1 - 2 ug/ml DB: 1 ug/ml

• Tested Reactivity: Human, Wide Range Predicted
• Host / Isotype: Mouse / IgG2a
• Class: Recombinant
• Type: Antibody
• Modification: Methylated
• Immunogen: This antibody was raised against a synthetic peptide containing trimethyl Lys4 of human histone H3.
• Full Name: AbFlex Histone H3K4me3 antibody (rAb)
• Synonyms: Histone H3, histone methylation, transcriptional activation chromatin, antibody, antibodies, monoclonal, H3K4me3, AbFlex, recombinant antibodies
• Molecular weight: 17 kDa
• GenBank accession number: NP_003522
• RRID: AB_2793824
• 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. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

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 H3K4me3 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). Histone H1 is a linker histone, present at the interface between the nucleosome core and DNA entry/exit points. Histone H1 is responsible for establishing higher-order chromatin structure. 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. Histone methylation can be associated with transcriptional activation or repression, depending on the methylated residue. Lysine 4 of histone H3 can be mono-, di- or trimethylated by different histone methyltransferases (HMTs) such as SET1 or ASH1. Methylation of Lys4 is often associated with transcriptional activation. The demethylase LSD1 is able to demethylate histone H3 Lys4.