Anti phospho s10 h3

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-phospho-S10 H3 is an antibody that specifically recognizes histone H3 phosphorylated at serine 10. This product can be used to detect and quantify the level of this histone modification in various biological samples.

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5 protocols using anti phospho s10 h3

Antibody Characterization for TAZ Signaling

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The TAZ (V386) antibody from Cell Signaling Technology was used for Western blotting throughout the study. Rabbit polyclonal phospho-specific antibodies against TAZ S90, S105, T326, and T346 were generated and purified by AbMart. Anti-β-actin, anti-GFP, and anti-cyclin B antibodies were from Santa Cruz Biotechnology. Anti-glutathione S-transferase (GST) and anti-His antibodies were from Bethyl Laboratories. Anti-phospho-S10 H3 and anti-vimentin antibodies were from Cell Signaling Technology. Anti-E-cadherin antibody was from BD Biosciences. Anti-β-tubulin (Sigma), anti-α-tubulin (Abcam), and anti-γ-tubulin (Biolegend) antibodies were used for immunofluorescence staining.

Zhang L., Chen X., Stauffer S., Yang S., Chen Y, & Dong J. (2015). CDK1 phosphorylation of TAZ in mitosis inhibits its oncogenic activity. Oncotarget, 6(31), 31399-31412.

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YAP Protein Immunoprecipitation and Western Blotting

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The YAP antibodies from Abnova (H00010413-M01) and Abcam (52771) were used for immunoprecipitation of endogenous YAP and for Western blotting, respectively, throughout the study. Rabbit polyclonal phospho-specific antibody against YAP T119 has been previously described [14 (link)]. Anti-β-actin, anti-Myc, and anti-cyclin B antibodies were from Santa Cruz Biotechnology. Anti-phospho-S10 H3 and anti-survivin antibodies were from Cell Signaling Technology. Immunoprecipitation and Western blotting assays were done as previously described [37 (link)].

Yang S., Zhang L., Purohit V., Shukla S.K., Chen X., Yu F., Fu K., Chen Y., Solheim J., Singh P.K., Song W, & Dong J. (2015). Active YAP promotes pancreatic cancer cell motility, invasion and tumorigenesis in a mitotic phosphorylation-dependent manner through LPAR3. Oncotarget, 6(34), 36019-36031.

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Antibody Characterization for PBK Phosphorylation

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The PBK and phospho-PBK (pT9) antibodies were purchased from Cell Signaling Technology (4942 and 4941). Rabbit polyclonal phospho-specific antibodies against PBK T24, S32, and S59 were generated and purified by AbMart. The peptides used for immunizing rabbits were SVLCS-pT-PTINI (T24), INIPA-pS-PFMQK (S32) and RGLSH-pS-PWAVK (S59). The corresponding non-phosphorylated peptides were also synthesized and used for antibody purification and blocking assays. Anti-Flag antibody was from Sigma. Anti-PLK1 was from BioLegend. Anti-β-actin, anti-Mps1/TTK, anti-cyclin E1, and anti-cyclin B antibodies were from Santa Cruz Biotechnology. Anti-aurora-A, anti-CDK2, anti-glutathione S-transferase (GST), anti-BUB1, and anti-BubR1 antibodies were from Bethyl Laboratories. Anti-phospho-S10 H3, anti-YAP, anti-phospho-S127 YAP, anti-phospho-S397 YAP, anti-vimentin, anti-E-cadherin, anti-N-cadherin, anti-CDC25C, anti-CDK4, anti-CDK5, anti-CDK6, anti-Cyclin A2, anti-Cyclin E2, anti-MAD2, anti-phospho-S795 Rb, anti-Wee1, and anti-phospho-S642 Wee1 antibodies were from Cell Signaling Technology. Anti-β-tubulin (Sigma) antibodies were used for immunofluorescence staining.

Stauffer S., Zeng Y., Zhou J., Chen X., Chen Y, & Dong J. (2017). CDK1-mediated Mitotic Phosphorylation of PBK is Involved in Cytokinesis and Inhibits its Oncogenic Activity. Cellular signalling, 39, 74-83.

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Cell Cycle Regulation and Apoptosis

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The fluorogenic probe 2′, 7′-dichlorofluorescin diacetate (DCFH-DA, D6883) was from Sigma. 5-bromo-2′-deoxyuridine (BrdU, 000103) was provided by Thermo Fisher Scientific. Anti-BrdU antibody (#5292S), anti-phospho-H3-S10 (#3377), anti-topoisomerase II α (#12286), anti-phospho-P53 (#9284), anti-P21(#2947), anti-CDK1(#9116), anti-phosphor-CDK1(Tyr15) (#9111), anti-Cyclin B1(#4138), anti-PARP (#9542) and anti-Caspase 3 (#9662) were purchased from Cell Signaling Technology, anti-P53 (#AP6266d) was from Abcepta. Anti-β-Tubulin and anti-GAPDH antibodies were from Beijing TransGen Biotech (Beijing, China). Prestained Protein Ladder (26616) and M-PER buffer were from Thermo Pierce. FITC Annexin V Apoptosis Detection Kit I (#556547) and PI/RNase Staining Buffer (550825) were from BD Biosciences, protease inhibitor and phosphatase inhibitor cocktails were from Roche and the PVDF membrane was from Millipore.

Yang X., Song C., Zhang L., Wang J., Yu X., Yu B., Zablotskii V, & Zhang X. (2021). An upward 9.4 T static magnetic field inhibits DNA synthesis and increases ROS-P53 to suppress lung cancer growth. Translational Oncology, 14(7), 101103.

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Protein Expression Analysis Protocol

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Verapamil, vincristine, and paclitaxel were purchased from Sigma-Aldrich (St. Louis, MO, USA). The commercial antibodies used included anti-β-actin (sc-47778, dilution of 1:10,000), anti-CDK1 (sc-54, dilution of 1:1000), anti-cyclin B1 (sc-752, dilution of 1:1000), and α-tubulin (sc-23948, dilution of 1:5000) antibodies purchased from Santa Cruz Biotechnology (Dallas, TX, USA); anti-phospho-H3-S10 (#9701, dilution of 1:1000), anti-phospho-CDK1-Y15 (#4539, dilution of 1:1000), anti-CDK2 (#2546, dilution of 1:1000), anti-CDK4 (#12790, dilution of 1:1000), and anti-H3 (#9715, dilution of 1:5000) antibodies purchased from Cell Signaling Technology (Danvers, MA, USA); anti-INCENP (ab12183, dilution of 1:1000), anti-survivin (ab76424, dilution of 1:1000), anti-aurora B (ab2254, dilution of 1:1000), and anti-phospho-H3-T3 (ab78351, dilution of 1:1000) antibodies purchased from Abcam (Cambridge, UK); and anti-borealin (NBP1-89951, dilution of 1:1000) antibody purchased from Novus Biologicals (Centennial, CO, USA). HRP-conjugated secondary antibodies (dilution of 1:10,000) were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA).

Park M., Hwang J.W., Cho Y., Kim S., Han S.H., Yu J., Ha S., Kim W.Y., Kim S.N., Kim I.S, & Kim Y.K. (2021). A novel synthetic microtubule inhibitor exerts antiproliferative effects in multidrug resistant cancer cells and cancer stem cells. Scientific Reports, 11, 10822.

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