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Antibodies

AbFlex® Histone H4K20me3 antibody (rAb) (biotin)

Catalog No: 91105 Format: 100 µg $405 Buy Now
Catalog No: 91106 Format: 10 µg $85 Buy Now

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

Applications

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

for AbFlex Histone H4K20me3 antibody (rAb) (biotin)Application Notes


Validated Applications:
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.

for AbFlex Histone H4K20me3 antibody (rAb) (biotin)Immunogen

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

for AbFlex Histone H4K20me3 antibody (rAb) (biotin)Buffer

Purified IgG in 50mM sodium phosphate pH 8.0, 150mM NaCl, and 100mM imidazole with 30% glycerol and 0.035% sodium azide.

AbFlex™ recombinant antibodies are genetically derived from DNA sequences of parental hybridoma clones.  For details on the parental clone, see Catalog No. 39671.

 
AbFlex<sup>®</sup> Histone H4K20me3 antibody (rAb) (biotin) tested by ChIP-Seq.

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.

AbFlex<sup>®</sup> Histone H4K20me3 antibody (rAb) (biotin) tested by immunofluorescence.

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<sup>®</sup> Histone H4K20me3 antibody (rAb) (biotin) tested by Western blot.

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<sup>®</sup> Histone H4K20me3 antibody (rAb) (biotin) tested by ELISA.

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.

for AbFlex Histone H4K20me3 antibody (rAb) (biotin)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, an avidin tag sequence 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 (biotin) was expressed as full-length IgG with mouse immunoglobulin heavy and light chains (IgG2a isotype) in mammalian 293 cells. The antibody was directly labeled with biotin using the biotin ligase, BirA.

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). 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 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.

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) (biotin).

Bridging Antibody to Improve ChIP, MeDIP and IP

Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use AbFlex® Histone H4K20me3 antibody (rAb) (biotin).

Compatible Secondary Antibodies

Active Motif has developed a variety of high-quality secondary antibodies for most applications, with anti-rabbit and anti-mouse conjugates to a broad line of high-quality fluorescent molecules, as well as horse radish peroxidase (HRP). Visit our Secondary Antibody Conjugates page to find the perfect secondaries for immunofluorescence experiments with AbFlex® Histone H4K20me3 antibody (rAb) (biotin).

for AbFlex Histone H4K20me3 antibody (rAb) (biotin)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.

for AbFlex Histone H4K20me3 antibody (rAb) (biotin)Guarantee

This product is guaranteed for 6 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;
  • 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;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting
 

Technical Data Sheet

AbFlex® Histone H4K20me3 antibody (rAb) (biotin)
 

Data Thumbnails

AbFlex<sup>®</sup> Histone H4K20me3 antibody (rAb) (biotin) tested by ChIP-Seq.

ChIP-Seq of AbFlex® Histone H4K20me3 antibody (rAb) (biotin).
(Click image to enlarge and see details.)

AbFlex<sup>®</sup> Histone H4K20me3 antibody (rAb) (biotin) tested by immunofluorescence.

Immunofluorescence stain of AbFlex® Histone H4K20me3 antibody (rAb) (biotin).
(Click image to enlarge and see details.)

AbFlex<sup>®</sup> Histone H4K20me3 antibody (rAb) (biotin) tested by Western blot.

Western blot of AbFlex® Histone H4K20me3 antibody (rAb) (biotin).
(Click image to enlarge and see details.)

AbFlex<sup>®</sup> Histone H4K20me3 antibody (rAb) (biotin) tested by ELISA.

ELISA of AbFlex® Histone H4K20me3 antibody (rAb) (biotin).
(Click image to enlarge and see details.)