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Ag014699

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AG014699 is a laboratory equipment item. It is a precision instrument designed for laboratory use. The core function of this product is to perform specific measurements and tasks required in a laboratory setting.

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Lab products found in correlation

Tubulin, by Merck Group (3 mentions) Rucaparib, by Chemietek (3 mentions) Parp1, by Cell Signaling Technology (3 mentions) Veliparib, by Chemietek (2 mentions) Par 4335 mc 100 ac, by Bio-Techne (2 mentions) Suz12, by Cell Signaling Technology (2 mentions) Gst ezh2, by Abcam (2 mentions) γh2ax, by Merck Group (2 mentions) Niraparib, by Chemietek (2 mentions) Olaparib, by Chemietek (2 mentions) Mk4827, by Chemietek (2 mentions) Erlotinib, by Selleck Chemicals (1 mentions) Gefitinib, by Selleck Chemicals (1 mentions) Sodium arsenite solution, by Merck Group (1 mentions) Cleaved caspase 3, by Abcam (1 mentions) Phosphorylated met, by Cell Signaling Technology (1 mentions) γ h2ax, by Abcam (1 mentions) Abt 888, by Chemietek (1 mentions) Flag tag (flag), by Merck Group (1 mentions) Parp 1, by Santa Cruz Biotechnology (1 mentions)

5 protocols using ag014699

1

Antibodies and Inhibitors for Cancer Research

Cited 1 time
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Antibodies used in this study are as follows: tubulin (#T5168) and Flag (#F3165) were purchased from Sigma-Aldrich (St. Louis, MO); PARP1 (#sc-7150) for Western blotting from Santa Cruz Biotechnology (Santa Cruz, CA); PARP1 (#9532) for immunoprecipitation (IP), MET (#8198), phosphorylated MET (#3077), phosphorylated EGFR for IHC (#3777) and EGFR (#4267) from Cell Signaling Technology (Danvers, MA); and phosphorylated EGFR for Western blotting (#5650), Ki67 (#21700), cleaved caspase 3 (#2302), γ-H2AX (#140498), and PAR (#14460) from Abcam (Cambridge, MA); 8-hydroxy-2′-deoxy guanosine (8-OHdG) from Genox Corporation (Baltimore, MD). The mouse phospho-Y907-PARP1 antibody was generated against a phosphorylated synthetic peptide (ADMVSKSAN-Yp-CHTSQGD) at China Medical University as described previously (13 (link)). The following inhibitors were used in this study: MET kinase inhibitor crizotinib (#C-7900) was purchased from LC Laboratories (Woburn, MA); EGFR inhibitor gefitinib (#S1025) and erlotinib (#S1023) from Selleck Chemicals (Houston, TX); PARP inhibitors ABT-888 (veliparib, #CT-A888) and AG014699 (rucaparib, #CT-AG01) from ChemieTek (Indianapolis, IN). Hydrogen peroxide (#216763) and sodium arsenite solution (#35000) were obtained from Sigma-Aldrich.
Dong Q., Du Y., Li H., Liu C., Wei Y., Chen M.K., Zhao X., Chu Y.Y., Qiu Y., Qin L., Yamaguchi H, & Hung M.C. (2018). EGFR and c-MET cooperate to enhance resistance to PARP inhibitors in hepatocellular carcinoma. Cancer research, 79(4), 819-829.
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2

Examining EZH2 and PARylation Interactions

Cited 2 times
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Expression of EZH2 protein and shRNA against EZH2 were described previously61 (link). Point mutations were introduced by site-directed mutagenesis as described previously16 (link). EED shRNA expression plasmids and siRNAs against SUZ12 were obtained from Thermo Fisher Scientific. The following antibodies were used: EZH2 (#5246, Cell Signaling Technology; 612667, BD Biosciences), EED (sc-28701, Santa Cruz Biotechnology; 09-774, EMD Millipore), SUZ12 (#3737, Cell Signaling Technology), PAR (4335-MC-100-AC, Trevigen; 551813, BD Biosciences), PARP1 (#9532, Cell Signaling Technology; sc-7150, Santa Cruz Biotechnology), HA (11666606001, Sigma-Aldrich; sc-805, Santa Cruz Biotechnology), Histone H3 (ab1791, Abcam; #3638, Cell Signaling Technology), H3-K27-me3 (ab6002, ab108245, Abcam), GST (sc-53909, Santa Cruz Biotechnology), γ-H2AX (#05-636, EMD Millipore), Tubulin (T5168, Sigma-Aldrich). Purified PARP protein and GST-EZH2 were purchased from Abcam and BPS Bioscience, respectively. Olaparib, EPZ-6438, GSK132 were purchased from Shelleck Chemicals, and AG014699 from ChemieTek.
Yamaguchi H., Du Y., Nakai K., Ding M., Chang S.S., Hsu J.L., Yao J., Wei Y., Nie L., Jiao S., Chang W.C., Chen C.H., Yu Y., Hortobagyi G.N, & Hung M.C. (2017). EZH2 contributes to the response to PARP inhibitors through its PARP-mediated poly-ADP ribosylation in breast cancer. Oncogene, 37(2), 208-217.
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3

Examining EZH2 and PARylation Interactions

Cited 2 times
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Expression of EZH2 protein and shRNA against EZH2 were described previously61 (link). Point mutations were introduced by site-directed mutagenesis as described previously16 (link). EED shRNA expression plasmids and siRNAs against SUZ12 were obtained from Thermo Fisher Scientific. The following antibodies were used: EZH2 (#5246, Cell Signaling Technology; 612667, BD Biosciences), EED (sc-28701, Santa Cruz Biotechnology; 09-774, EMD Millipore), SUZ12 (#3737, Cell Signaling Technology), PAR (4335-MC-100-AC, Trevigen; 551813, BD Biosciences), PARP1 (#9532, Cell Signaling Technology; sc-7150, Santa Cruz Biotechnology), HA (11666606001, Sigma-Aldrich; sc-805, Santa Cruz Biotechnology), Histone H3 (ab1791, Abcam; #3638, Cell Signaling Technology), H3-K27-me3 (ab6002, ab108245, Abcam), GST (sc-53909, Santa Cruz Biotechnology), γ-H2AX (#05-636, EMD Millipore), Tubulin (T5168, Sigma-Aldrich). Purified PARP protein and GST-EZH2 were purchased from Abcam and BPS Bioscience, respectively. Olaparib, EPZ-6438, GSK132 were purchased from Shelleck Chemicals, and AG014699 from ChemieTek.
Yamaguchi H., Du Y., Nakai K., Ding M., Chang S.S., Hsu J.L., Yao J., Wei Y., Nie L., Jiao S., Chang W.C., Chen C.H., Yu Y., Hortobagyi G.N, & Hung M.C. (2017). EZH2 contributes to the response to PARP inhibitors through its PARP-mediated poly-ADP ribosylation in breast cancer. Oncogene, 37(2), 208-217.
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4

Synthesis and Storage of PARP Inhibitors

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Olaparib (AZD2281; Chemietek), niraparib (MK4827; Chemietek), rucaparib (AG014699; Chemietek), and AZD5363 (Active Biochem) were dissolved in DMSO at a concentration of 10 mM and stored at −20 °C. 6-alkyne (6Yn)- and 2-alkyne (2Yn)-adenosine probes were synthesized in-house by the chemistry unit of the Chemical Biology Core of the Moffitt Cancer Center. Drug dilutions were made in DMSO.
Deng O., Dash S., Nepomuceno T.C., Fang B., Yun S.Y., Welsh E.A., Lawrence H.R., Marchion D., Koomen J.M., Monteiro A.N, & Rix U. (2022). Integrated proteomics identifies PARP inhibitor-induced prosurvival signaling changes as potential vulnerabilities in ovarian cancer. The Journal of Biological Chemistry, 298(11), 102550.
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5

PARP Inhibitor Preparation Protocol

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Olaparib (AZD2281; Chemietek), niraparib (MK4827; Chemietek), rucaparib (AG014699; Chemietek), Veliparib (CT-A888; Chemietek), talazoparib (BMN673; Cayman Chemical and Moffitt Cancer Center) and adavosertib (MK1775; Chemietek) were dissolved in DMSO at a concentration of 10 mM and stored at −20°C. (Rac)-i-Talazoparib (2) was made in house using the chemistry unit of the Chemical Biology Core of the Moffitt Cancer Center. Drug dilutions were made in DMSO as necessary.
Palve V., Knezevic C.E., Bejan D.S., Luo Y., Li X., Novakova S., Welsh E.A., Fang B., Kinose F., Haura E.B., Monteiro A.N., Koomen J.M., Cohen M.S., Lawrence H.R, & Rix U. (2021). The non-canonical target PARP16 contributes to polypharmacology of the PARP inhibitor talazoparib and its synergy with WEE1 inhibitors. Cell chemical biology, 29(2), 202-214.e7.
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