ChIP analysis of sheared chromatin from 1x10⁵ K562 cells using GTX60828 Histone H4K20me3 (trimethyl Lys20) antibody - ChIP grade. The IP'd DNA was analysed by QPCR as described above (figure 2A). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2B shows the signal distribution along the long arm of chromosome 19 and a zoomin to an enriched region containing several ZNF repeat genes. Figure 2C and D show the enrichment in the telomeric region of chromosome 12, also containing several ZNF repeat genes, and at ZNF510 on chromosome 9, respectively. The position of the amplicon used for ChIP-qPCR is indicated by an arrow.
Antibody amount : 0.5μg

ELISA analysis of peptide containing the histone modification of interest using GTX60828 Histone H4K20me3 (trimethyl Lys20) antibody - ChIP grade.

ChIP analysis of sheared chromatin from 10⁶ HeLa cells using GTX60828 Histone H4K20me3 (trimethyl Lys20) antibody - ChIP grade. The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration consisting of 0.2, 0.5, 1 and 2 μg of antibody per ChIP experiment was analyzed. IgG (1 μg/IP) was used as a negative IP control. Figure 1A. Quantitative PCR was performed with primers specific for the promoter of the active GAPDH and EIF4A2 genes, used as negative controls, and for the ZNF10 gene and the Sat2 satellite repeat, used as positive controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B. Recovery of the nucleosomes carrying the H4K20me1, H4K20me2, H4K2me3 and the unmodified H4K20 as determined by qPCR. The figure clearly shows the antibody is very specific in ChIP for the H4K20me3 modification.

WB analysis of whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using GTX60828 Histone H4K20me3 (trimethyl Lys20) antibody - ChIP grade.
Dilution : 1:1,000

Protein Array analysis of an array containing 384 peptides with different combinations of modifications from histone H3, H4, H2A and H2B using GTX60828 Histone H4K20me3 (trimethyl Lys20) antibody - ChIP grade.This figure shows the specificity factor, calculated as the ratio of the average intensity of all spots containing the mark, divided by the average intensity of all spots not containing the mark.
Dilution : 1:10,000

ICC/IF analysis of methanol fixed HeLa cells using GTX60828 Histone H4K20me3 (trimethyl Lys20) antibody - ChIP grade.
Green : Primary antibody
Blue : DAPI
Dilution : 1:500

Dot blot analysis of 0.2 - 100 pmol of the peptides containing other histone modifications and the unmodified H4K20 using GTX60828 Histone H4K20me3 (trimethyl Lys20) antibody - ChIP grade.
Dilution : 1:20,000