Histone H4K20me3 antibody (mAb)

RRID: AB_2650526
Clone: 6F8-D9
Catalog No: 39671 Format: 100 µg $455 Buy
Catalog No: 39672 Format: 10 µg $110 Buy

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Antibody Type:
Protein G Chromatography
Molecular Weight:
8 kDa
Human, Mouse, Wide Range Predicted


ChIP Validated Western Blot Validated Dot Blot Validated Immunocytochemistry Validated

Application Notes

Validated Applications:
ChIP: 5 - 10 µg per ChIP
WB: 1 - 2 µg/ml dilution
DB: 0.2 µg/ml dilution

Slight reactivity towards histone H4 dimethyl Lys20 might be observed under certain conditions. Individual optimization may be required.

Published Applications:

*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.

The addition of 0.05% Tween 20 in the blocking buffer and primary antibody incubation buffer can be used to aid in detection by Western blot. Individual optimization may be required.


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


Purified IgG in PBS with 0.035% sodium azide. Sodium azide is highly toxic.

View our Guide to Histone Modifications and Biological Function.



Histone H4 trimethyl Lys20 mAb (Clone 6F8-D9) tested by ChIP.
Chromatin IP performed using the ChIP-IT® Express Kit (Catalog No. 53008) and HeLa chromatin (1.5 x 106 cell equivalents per ChIP) using 5 µg of Histone H4 trimethyl Lys20 antibody (mAb) (Clone 6F8-D9) or the equivalent amount of rabbit IgG as a negative control. Real time, quantitative PCR (RT-qPCR) was performed on DNA purified from each of the ChIP reactions using a primer pair specific for human SAT-alpha repeats. Data are presented as Fold Enrichment of the ChIP antibody signal compared to the negative control IgG (which has been normalized to 1.0) using the ddCT method.

Histone H4 trimethyl Lys20 antibody (mAb) (Clone 6F8-D9) tested by Western blot.
HeLa nuclear extract (20 µg per lane) probed with Histone H4 trimethyl Lys20 antibody (mAb) (Clone 6F8-D9) (2 µg/ml dilution).

Histone H4 trimethyl Lys20 mAb (Clone 6F8-D9) tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone Histone H4 trimethyl Lys20 antibody (mAb) (Clone 6F8-D9) for trimethyl-Lys20 of histone H4. Modified and unmodified peptides were spotted onto PVDF and probed with Histone H4 trimethyl Lys20 antibody (mAb) (Clone 6F8-D9) at a dilution of 2 µg/ml. The amount of peptide (in picomoles) spotted is indicated next to each row.

Lane 1: monomethyl Lys20 peptide.
Lane 2: dimethyl Lys20 peptide.
Lane 3: trimethyl Lys20 peptide.
Lane 4: Unmodified Lys20 peptide.


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 Histone H4K20me3 antibody (mAb).

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 Histone H4K20me3 antibody (mAb).


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.


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;
  • 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