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H3k79me3

Manufactured by Cell Signaling Technology
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H3K79me3 is a product that detects trimethylation of lysine 79 on histone H3, which is a post-translational modification associated with active transcription. This antibody can be used in various applications, including chromatin immunoprecipitation (ChIP), western blotting, and immunofluorescence.

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Lab products found in correlation

H3k36me3, by Cell Signaling Technology (4 mentions) H3k36me2, by Cell Signaling Technology (3 mentions) H3k9me2, by Cell Signaling Technology (3 mentions) H3k9me3, by Cell Signaling Technology (3 mentions) H3k27me3, by Cell Signaling Technology (3 mentions) H3k4me3, by Cell Signaling Technology (3 mentions) Ab113476, by Abcam (2 mentions) β actin, by Merck Group (2 mentions) Western lightning plus ecl, by PerkinElmer (1 mentions) Pvdf membrane, by Bio-Rad (1 mentions) H4k20me3, by Cell Signaling Technology (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) Cleaved parp, by Cell Signaling Technology (1 mentions) Dot1l, by Cell Signaling Technology (1 mentions) Annexin 5 apoptosis detection kit, by Merck Group (1 mentions) Flag tag (flag), by Cell Signaling Technology (1 mentions) Hoxa9, by Abcam (1 mentions) H3k4me2, by Cell Signaling Technology (1 mentions) H3k27me2, by Cell Signaling Technology (1 mentions) Npm1 wt, by Thermo Fisher Scientific (1 mentions)

6 protocols using h3k79me3

1

Western Blot Analysis of Histone Modifications

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Tumors were crushed with potter, and lysed in a RIPA lysis buffer (Tris-HCl 50 mM pH 7.5; IGEPAL 1%; NaCl 150 mM; EGTA 1 mM; NaF 1 mM) supplemented with phosphatase inhibitors (NA3VO4 10 µL/mL) and protease inhibitors (leupeptin 1 µL/mL; aprotinin 1 µL/mL; pepstatin 1 µL/mL; and phenylmethylsulfonyl fluoride 4 µL/mL). Protein extracts were migrated in SDS-PAGE, and transferred to polyvinylidene difluoride (PVDF) membranes (Bio-Rad, Marnes-la-Coquette, France). After probing with primary antibodies, the membranes were incubated with horseradish peroxidase-conjugated secondary antibodies, and signals visualized by Western Lightning® Plus-ECL (Perkin Elmer, Villebon S/Yvette, France). Antibodies specific for PARP (#5246), H3K4me3 (#9727), H3K9me3 (#13969), H3K27me3 (#9733), H3K36me3 (4909), H3K79me3 (#4260), H4K20me3 (#5737), and H3 (#4499) were obtained from Cell Signalling (Danvers, MA, USA), and actin (sc47778) was provided by Santa Cruz (Heidelberg, Germany). The whole Western blot figures can be found in File S1.
Lhuissier E., Aury-Landas J., Lenté M., Boumediene K, & Baugé C. (2021). Co-Treatment with the Epigenetic Drug, 3-Deazaneplanocin A (DZNep) and Cisplatin after DZNep Priming Enhances the Response to Platinum-Based Therapy in Chondrosarcomas. Cancers, 13(18), 4648.
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2

Histone Modifications Profiling Protocol

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Histones were isolated with a histone extraction kit (ab113476; Abcam), following the manufacturer’s procedure. Histone Protein (2 μg) was separated by 4–20% Tris-Glycine gels, and transferred to nitrocellulose membranes. Primary antibodies used for Western blotting are as follows: H3K4Me3, H3K9Me2, H3K9Me3, H3K27Me3, H3K36Me2, H3K36Me3, H3K79Me3 (Cell Signaling biotechnology, 1:1000).
Wei S., Wang J., Oyinlade O., Ma D., Wang S., Kratz L., Lal B., Xu Q., Liu S., Shah S.R., Zhang H., Li Y., Quiñones-Hinojosa A., Zhu H., Huang Z.Y., Cheng L., Qian J, & Xia S. (2018). Heterozygous IDH1R132H/WT created by “single base editing” inhibits human astroglial cell growth by downregulating YAP. Oncogene, 37(38), 5160-5174.
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3

Antibodies for Protein Analysis Techniques

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The following primary antibodies were used in western blotting assays: NPM1 mutant (NB110-61646, Novus, Littleton, USA), NPM1 WT (Thermo scientific, Rockford, USA), NPM1 (Sigma-Aldrich, St. Louis, USA), PBX3 (Abcam, Cambridge, USA), HOXA9 (Abcam, Cambridge, USA), Cleaved caspase 3, Cleaved PARP (Cell Signaling, Danvers, USA), Flag, H3K4me2, H3K9me2, H3K27me2, H3K36me2, H3K79me1, H3K79me2, H3K79me3, H3, DOT1L (Cell Signaling, Danvers, USA), and β-Actin (Sigma, St. Louis, MO, USA).
All antibodies and the following kits applied in flow cytometry analyses were purchased from BD Biosciences (BD Pharmingen™, New Jersey, USA): CD45.1, CD45.2, Mac-1, Gr-1, Sca-1, c-Kit, Mouse Lineage Panel, AnnexinV Apoptosis Detection kit, and BrdU Flow kit.
Immunofluorescence staining was performed with Flag (Cell Signaling, Danvers, USA), H3K79me2 (Cell Signaling, Danvers, USA and PMT-bio, Hangzhou, China), PBX3 (Santa Cruz, California, USA), HOXA9 (Abcam, Cambridge, USA) and NPM1 antibodies as following: NPM1 mutant (Novus, Littleton, USA), NPM1 WT (Thermo scientific, Rockford, USA), NPM1 (Cell Signaling, Danvers, USA).
Zhang W., Zhao C., Zhao J., Zhu Y., Weng X., Chen Q., Sun H., Mi J.Q., Li J., Zhu J., Chen Z., Pandolfi P.P., Chen S., Yan X, & Xu J. (2018). Inactivation of PBX3 and HOXA9 by down-regulating H3K79 methylation represses NPM1-mutated leukemic cell survival. Theranostics, 8(16), 4359-4371.
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4

Chromatin Profiling of Histone Modifications

Cited 1 time
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Chromatin immunoprecipitation was performed as described previously [27 (link)] 48 h after transfection using antibodies against H2Bub1 (Cat. No. 5546S, Cell Signaling Technology), H3K79me3 (Cat. No. C15410068, Diagenode), H3K27ac (Cat. No. C15410196, Diagenode) and RNApol-II (Cat. No. C15200004, Diagenode). Next-generation sequencing library was prepared using the KAPA Hyper Prep Kit (KR0961–v6.17) according to manufacturer’s instructions and samples were sequenced (single-end 50 bp) on a HiSeq4000 (Illumina) at the NGS Integrative Genomics Core Unit (NIG) at the University Medical Center Göttingen (ArrayExpress accession: E-MTAB-12000). Processing of sequencing data was performed in the Galaxy environment (galaxy.gwdg.de). Briefly, ChIP-seq reads were mapped to the hg19 reference genome assembly using Bowtie2 (version 2.3.4.2). PCR duplicates were removed using the RmDup tool (version 2.0.1). The bamCoverage tool (version 3.2.0.0.0) was utilized to generate normalized coverage files using the 1x depth (reads per genome coverage, RPGC) as a normalizing method. Peak calling was performed using the MACS2 callpeak (version 2.1.1.20160309.0), and computeMatrix and plotHeatmap (version 2.5.1.1.0) to generate aggregate plots and heatmaps, respectively. Occupancy profiles were visualized using the Integrative Genomics Viewer (IGV 2.8.0).
Prokakis E., Jansari S., Boshnakovska A., Wiese M., Kusch K., Kramm C., Dullin C., Rehling P., Glatzel M., Pantel K., Wikman H., Johnsen S.A., Gallwas J, & Wegwitz F. (2023). RNF40 epigenetically modulates glycolysis to support the aggressiveness of basal-like breast cancer. Cell Death & Disease, 14(9), 641.
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5

Histone Modifications Profiling Protocol

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Histones were isolated with a histone extraction kit (ab113476; Abcam), following the manufacturer’s procedure. Histone Protein (2 μg) was separated by 4–20% Tris-Glycine gels, and transferred to nitrocellulose membranes. Primary antibodies used for Western blotting are as follows: H3K4Me3, H3K9Me2, H3K9Me3, H3K27Me3, H3K36Me2, H3K36Me3, H3K79Me3 (Cell Signaling biotechnology, 1:1000).
Wei S., Wang J., Oyinlade O., Ma D., Wang S., Kratz L., Lal B., Xu Q., Liu S., Shah S.R., Zhang H., Li Y., Quiñones-Hinojosa A., Zhu H., Huang Z.Y., Cheng L., Qian J, & Xia S. (2018). Heterozygous IDH1R132H/WT created by “single base editing” inhibits human astroglial cell growth by downregulating YAP. Oncogene, 37(38), 5160-5174.
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6

Histone Methylation Analysis in Osteoclastogenesis

Cited 2 times
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Total cell lysates and nuclear protein-enriched lysates were collected using appropriate cell lysis buffers, and western blot analysis was performed as described previously40 (link),48 (link). Analysis of histone methylation was performed 24 h after induction of osteoclastogenesis, unless otherwise specified. Proteins were separated by SDS–PAGE, transferred to nitrocellulose membranes and incubated overnight with primary antibodies against β-actin (A5441, Sigma-Aldrich), c-MYC (5605, Cell Signaling Technology), H3K4Me3 (9751, Cell Signaling Technology), H3K9Me3 (13969, Cell Signaling Technology), H3K27Me3 (9733, Cell Signaling Technology), H3K36Me3 (4909, Cell Signaling Technology), H3K79Me3 (74073, Cell Signaling Technologies), lamin A/C (sc-376248, Santa Cruz Biotechnologies), NFATc1 (sc-7294, Santa Cruz Biotechnologies), PHGDH (66350, Cell Signaling Technologies) or PSAT1 (NBP1-55368, Bio-Techne). Appropriate anti-mouse or anti-rabbit horseradish peroxidase-conjugated secondary antibodies were used for chemiluminescent protein detection (Western Lightning Plus, PerkinElmer).
Stegen S., Moermans K., Stockmans I., Thienpont B, & Carmeliet G. (2024). The serine synthesis pathway drives osteoclast differentiation through epigenetic regulation of NFATc1 expression. Nature Metabolism, 6(1), 141-152.
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