AbFlex Histone H4K20me3 antibody (rAb)

Host / Isotype

Mouse / IgG2a

Reactivity

Human, Wide Range Predicted

Applications

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

Cat No : 91107,91108

Synonyms

histone H4, histone, H4, H4K20me3, Histone H4 trimethyl Lys20, Histone H4 trimethyl Lysine 20, Histone H4 trimethyl Lys 20, trimethyl, Lys20, K20, K20me3, me3, antibody, antibodies, 6F8-D9, H4K20Me3, chromatin immunoprecipitation, chip, recombinant, rAb, rec Ab, AbFlex, 6F8-D9

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Validation Data Gallery

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  AbFlex Histone H4K20me3 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 T-47D, mammary epithelial ductal cells and 4 ug of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 20 million sequence tags were mapped to identify Histone H4K20me3 binding sites. The image shows binding across a region of chromosome 19.

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  AbFlex H4K20me3 antibody (rAb) tested by immunofluorescence. HeLa cell stained with 2ug /mL of AbFlex H4K20me3 antibody (rAb) followed by anti-mouse-IgG-488.

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  AbFlex Histone H4K20me3 antibody (rAb) tested by Western blot. HeLa nuclear extract (20 ug per lane) probed with AbFlex Histone H4K20me3 antibody (rAb) ) (2 ug/ml dilution).

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  AbFlex Histone H4K20me3 antibody (rAb) tested by Luminex bead-based specificity analysis. Luminex bead-based specificity analysis was used to confirm the specificity of AbFlex Histone H4K20me3 antibody (rAb) antibody for Histone H4K20me3. Various methyl-lysine H4 or H3 peptides were conjugated to MagPlex Luminex beads and incubated with various amounts of AbFlex Histone H4K20me3antibody (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.

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Product Information

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

  Applications Validated by Active Motif: ChIP-Seq: 4 ug per ChIP ICC/IF: 1 - 2 ug/ml WB*: 0.5 - 2 ug/ml dilution Bead-based ELISA: 6 - 164 ng/ml Slight reactivity towards histone H4 dimethyl Lys20 might be observed under certain conditions. Individual optimization may be required. AbFlex recombinant antibodies are genetically derived from DNA sequences of parental hybridoma clones. For details on the parental clone, see Catalog No. 39671. *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 Histone H4 trimethyl Lys20 antibody (rAb) was raised against a peptide containing trimethyl Lys20 of Histone H4.
• Full Name: AbFlex Histone H4K20me3 antibody (rAb)
• Synonyms: histone H4, histone, H4, H4K20me3, Histone H4 trimethyl Lys20, Histone H4 trimethyl Lysine 20, Histone H4 trimethyl Lys 20, trimethyl, Lys20, K20, K20me3, me3, antibody, antibodies, 6F8-D9, H4K20Me3, chromatin immunoprecipitation, chip, recombinant, rAb, rec Ab, AbFlex, 6F8-D9
• Molecular weight: 8 kDa
• GenBank accession number: NP_778224
• RRID: AB_2793777
• 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 H4K20me3 antibody was expressed as full-length IgG with mouse immunoglobulin heavy and light chains (IgG2a isotype) in mammalian 293 cells. Histone H4 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; it 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; they play a major role in regulating gene expression. Histone 4 lysine 20 (H4K20) can be mono-, di- or trimethylated by different histone methyltransferases such as NSD1 or ASH1. The methylation of this lysine is often associated with transcriptional repression.