AbFlex® Histone H4K20me3 antibody (rAb)

RRID: AB_2793777
Catalog No: 91107 Format: 100 µg ¥67,000 Buy
Catalog No: 91108 Format: 10 µg ¥19,000 Buy

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Antibody Type:
Recombinant
Isotype:
IgG2a
Purification:
Ni-NTA
Host:
Mouse
Molecular Weight:
8 kDa
Reactivity:
Human, Wide Range Predicted

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Immunofluorescence Validated Enzyme-linked Immunosorbent Assay Validated Immunocytochemistry Validated

Application Notes

Applications Validated by Active Motif:
ChIP-Seq: 4 µg per ChIP
ICC/IF: 1 - 2 µg/ml
WB*: 0.5 - 2 µg/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.

Immunogen

This Histone H4 trimethyl Lys20 antibody (rAb) was raised against a peptide containing trimethyl Lys20 of Histone H4.

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.

View our Guide to Histone Modifications and Biological Function.

 

AbFlex® Histone H4K20me3 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 T-47D, mammary epithelial ductal cells and 4 µg 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. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

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

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

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.

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 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.

Positive Control

Active Motif's HeLa acid extract (Catalog No. 36200) can be used as a positive control for Histone H4 trimethyl Lys20 antibody.

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 H4K20me3 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