Histone H4K20me3 antibody (pAb) Catalog No: 39180, 39181 Purification: None Isotype: Serum Host: Rabbit Application(s): ChIP, IF, IP, WB Volume: 200 µl, 10 µl Reactivity: Human, Other (Wide Range) Molecular Weight: 8 kDa

Background: H4 is one of the core components of the . The nucleosome is the smallest subunit of 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 ). Chromatin is subject to a variety of chemical modifications, including post- translational modifications of the histone proteins and the 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 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.

Immunogen: This Histone H4 trimethyl Lys20 antibody was raised against peptide including trimethyl-Lys20 of human histone H4.

Buffer: Rabbit serum containing 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

Application Notes: This Histone H4 trimethyl Lys20 antibody (H4K20me) has been validated for use in Western blotting (1:1,000 - 1:5,000 dilution), chromatin IP (10 µl per ChIP) and reported to work in immunofluorescence; its specificity was verified by peptide competition. See References for additional details.

References: This antibody is described and used in Sims, J.K. et al. (2006) J Biol Chem 281: 12760-12766.

Storage and Guarantee: Antibodies in solution can be stored at -20°C for 2 years. Avoid repeated freeze/thaw cycles and keep on ice when not in storage. 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; FACS = Flow Cytometry; IF = Immunofluorescence; IHC = Immunohistochemistry; IP = Immunoprecipitation; WB = Western Blot North America 877 222 9543 • Europe +32 (0)2 653 0001 • Japan +81 (0)3 5225 3638 • www.activemotif.com Histone H4 trimethyl Lys20 antibody tested by Western blot. Detection of trimethylated Histone H4 by Western blot. The analysis was performed using HeLa acid extract and Histone H4 trimethyl Lys20 antibody at a 1:1,000 dilution.

Histone H4 trimethyl Lys20 antibody tested by ChIP.

® Chromatin IP performed using the ChIP-IT Express Kit (Catalog No. 53008) and HeLa 6 chromatin (1.5 x 10 cell equivalents per ChIP) using 10 µl of Histone H4 trimethyl Lys20 antibody 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 the indicated gene. 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.

North America 877 222 9543 • Europe +32 (0)2 653 0001 • Japan +81 (0)3 5225 3638 • www.activemotif.com