Plk1 thr210

Manufactured by Cell Signaling Technology

Sourced in United States

PLK1 (Thr210) is a protein that plays a critical role in cell division and mitosis. It is a phosphorylation site on the Polo-like kinase 1 (PLK1) protein, which is a key regulator of cell cycle progression. The phosphorylation of PLK1 at Thr210 is essential for its activation and proper localization during mitosis.

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), Phospho-PLK1 (Thr210) (4 citations) P-PLK1 (Thr210) (2 citations) Anti-phospho-PLK1 (Thr210), (2 citations)

3 protocols using plk1 thr210

Western Blotting for Cellular Signaling

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Western blotting was performed as previously described (10 (link)). We used the following primary antibodies: rabbit polyclonal antibody specific for PARP, phospho-TBK1/NAK (Ser¹⁷²), TBK1, phospho-polo-like kinase-1 (PLK1) (Thr²¹⁰), and PLK1from Cell Signaling (Beverly, MA), and β-actin antibody from Sigma (St. Louis, MO).

Li J., Fang B., Kinose F., Bai Y., Kim J.Y., Chen A., Rix U., Koomen J.M, & Haura E.B. (2016). Target identification in small cell lung cancer via integrated phenotypic screening and activity-based protein profiling. Molecular cancer therapeutics, 15(2), 334-342.

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Synthesis and Evaluation of HDAC6/Tubulin Inhibitor MPT0B451

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MPT0B451, a dual HDAC6 and tubulin inhibitor was synthesized by Dr. Jing-Ping Liou (School of Pharmacy, College of Pharmacy, Taipei Medical University, Taiwan). Vincristine, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and propidium iodide (PI) were purchased from Sigma Chemical Co. (St. Louis, MO, United States). Primary antibodies against Caspase 8 (06-775), MPM2 (pSer/pThr) (05-368) and GAPDH (AB2302) were purchased from Millipore (Bedford, MA, United States). The antibodies against Acetyl-Histone H3 (Lys9) (#9649), Histone H3 (#9715), Acetyl-α-tubulin (#3971), α-tubulin (#2144), Cyclin B1(#4135), Aurora A (#14475), Aurora A (Thr288) (#3079), PLK1 (#4535), PLK1 (Thr210) (#9062), cleaved caspase-3 (#9664), PARP (#9542), and caspase 9 (#9508) were purchased from Cell Signaling Technology (Beverly, MA, United States). The antibodies against Cdc2 (SC-54) and cdc25c (SC-372) as well as the labeled secondary antibodies goat anti-rabbit IgG-HRP (SC-2004) and goat anti-mouse IgG-HRP (SC-2005) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, United States).

Wu Y.W., Hsu K.C., Lee H.Y., Huang T.C., Lin T.E., Chen Y.L., Sung T.Y., Liou J.P., Hwang-Verslues W.W., Pan S.L, & HuangFu W.C. (2018). A Novel Dual HDAC6 and Tubulin Inhibitor, MPT0B451, Displays Anti-tumor Ability in Human Cancer Cells in Vitro and in Vivo. Frontiers in Pharmacology, 9, 205.

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Notch1-ICD Functional Analysis Protocol

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The following reagents were purchased from Santa Cruz Biotechnology: Fbxw7 and tubulin. In addition, we used Notch1 Val-1744, Notch1 D1E11, PLK1 208G4, and PLK1(Thr-210) from Cell Signaling Technology (Beverly, MA). γ-Secretase inhibitor IX (DAPT), was purchased from Calbiochem (Merck KGaA), dissolved in DMSO, and stored at −20 ºC until use. All cell extracts were prepared as described previously (30 (link)) and according to the manufacturer's instructions for detection of phospho-ERK (Cell Signaling Technology). The kinase library of 378 structurally diverse, cell-permeable kinase inhibitors was purchased from Selleckchem (Houston, TX) (catalogue no. L1200) (Table S1).
Notch1-ICD encodes the expression of human Notch1-IC from amino acid 1757 to 2555 and has been described previously (9 (link)). GST-NOTCH1-IC plasmid encodes the GST-Notch1-IC fusion protein encoding the mouse NOTCH1-IC region 1753–2531 was kindly provided by Dr. Lendhal (Karolinska Institute, Stockholm, Sweden) and described previously (31 (link)). The plasmids containing mutations in Notch1-ICD encoding the expression of human Notch1-IC from amino acid 1757 to 2555 were generated using the QuikChange II XL site-directed mutagenesis kit (Thermo Fisher Scientific) and verified by sequencing.

De Blasio C., Zonfrilli A., Franchitto M., Mariano G., Cialfi S., Verma N., Checquolo S., Bellavia D., Palermo R., Benelli D., Screpanti I, & Talora C. (2019). PLK1 targets NOTCH1 during DNA damage and mitotic progression. The Journal of Biological Chemistry, 294(47), 17941-17950.

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