Chromatin Immunoprecipitation (ChIP) was performed using commercial Protein-G coated 96 well high-throughput ChIP assay kit by loading 3 µg of cross-linked chromatin samples from Jurkat cells with either A) 1:50 dilution of Go-ChIP-Grade™ Purified anti-Histone H4 Monomethyl (Lys20) (clone 5E10-D8), B) equal amount of Purified Mouse IgG1 Isotype Control Antibody (clone MG1-45), or C) competitor's ChIP-grade Purified anti-Histone H4 Monomethyl (Lys20) Antibody and D) equal amount of matched Isotype Control Antibody as recommended by the manufacturer. The enriched DNA was purified and quantified by real-time qPCR using primers targeting human GAPDH gene region. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the 5% of total amount of input chromatin.

Total lysates (15µg protein) from Jurkat (lane1), Raw 264.7 (lane 2) and UMR106 (lane 3) cells were resolved by electrophoresis (4-20% Tris-Glycine gel), transferred to nitrocellulose, and probed with 1:5000 diluted (0.1 µg/mL) Purified anti-Histone H4 Monomethyl (Lys20) Antibody, clone 5E10-D8 (upper). Proteins were visualized by chemiluminescence detection using a 1:3000 diluted anti-Mouse-IgG secondary antibody conjugated to HRP for the anti-Histone H4 Monomethyl (Lys20) Antibody or 1:5000 diluted Direct-Blot™ HRP anti-β-Actin Antibody, clone 2F1-1(lower). Lane M: Molecular weight ladder.

IHC staining of purified anti-Histone H4 Monomethyl (Lys20) antibody (clone 5E10-D8) on formalin-fixed paraffin-embedded mouse brain tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., the tissue was incubated with 10 µg/ml of the primary antibody overnight at 4°C. BioLegend's Ultra-Streptavidin (USA) HRP kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective. Scale bar: 50 µm

IHC staining of purified anti-Histone H4 Monomethyl (Lys20) antibody (clone 5E10-D8) on formalin-fixed paraffin-embedded human brain tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., the tissue was incubated with 5 µg/ml of the primary antibody overnight at 4°C. BioLegend's Ultra-Streptavidin (USA) HRP kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective. Scale bar: 50 µm