Cp466722

Manufactured by Selleck Chemicals

Sourced in United States

CP466722 is a laboratory equipment item used for scientific research and analysis. It is a specialized device designed to perform specific functions in a laboratory setting. The core function of this product is to enable researchers to conduct their experiments and analyses effectively. No further details are provided to maintain an unbiased and factual approach.

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Lab products found in correlation

Lopac1280 library, by Merck Group (1 mentions) Etoposide, by Merck Group (1 mentions) Nutlin 3, by Merck Group (1 mentions) Ku55933, by Selleck Chemicals (1 mentions) Etoposide, by Corning (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Jak inhibitor 1, by Merck Group (1 mentions) Stattic, by Merck Group (1 mentions) Cwr22rv1, by American Type Culture Collection (1 mentions) C4 2 cells, by National Collection of Authenticated Cell Cultures (1 mentions) Transwell support, by Corning (1 mentions) Pd l1 antibody, by Thermo Fisher Scientific (1 mentions)

3 protocols using cp466722

High-Throughput Screening of Small Compounds

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The ATM inhibitors KU-55933 (S1092) and CP-466722 (S2245) were obtained from Selleckchem. LOPAC-1280 library, Nutlin-3 and Etoposide were purchased from Sigma-Aldrich.
Screening of the LOPAC small compound library was performed as follows: 10 000 cells per well were seeded in 96-well flat bottom plates (Corning CellBind CLS3340-50EA), 24 hours later cells were treated with drugs for 24 hours at a final concentration of 5μM in 0.5% DMSO in the presence of 0.5μM Etoposide, or with 0.5μM Etoposide in drug-free DMSO (negative control) or with DMSO only. Cells were then fixed for 10 minutes in 10% neutral buffered formalin solution (Sigma HT5012), nuclei stained for 10 minutes with DAPI (1μg/ml in PBS) and stored at 4°C until scanned (Thermo Cellomics Compartmental Analysis V4). Acquisitions were made with a 20x objective, both with XF93-Hoechst filter, 0.05sec exposure (DAPI) and XF93-TRITC filter, 0.5sec (RFP).

Mondesert O., Frongia C., Clayton O., Boizeau M.L., Lobjois V, & Ducommun B. (2015). Monitoring the Activation of the DNA Damage Response Pathway in a 3D Spheroid Model. PLoS ONE, 10(7), e0134411.

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Molecular Modulation of CRPC Cell Lines

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CRPC cell lines C4-2 and CWR22Rv1 were used in the present study. C4-2 cells were obtained from the China Center for Type Culture Collection (Wuhan, China). CWR22Rv1 cells were purchased from the American Type Culture Collection (Manassas, VA, USA). Cells were cultured in RPMI-1640 (Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% charcoal stripped fetal bovine serum (FBS; Thermo Fisher Scientific, Inc.). CP466722 (Selleck Chemicals, Houston, TX, USA), JAK inhibitor 1 (EMD Millipore, Billerica, MA, USA) and Stattic (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) inhibitors were added to cultured tumor cells (1:1,000 for all inhibitors) at 37°C, at 6, 12 and 24 h time intervals to inhibit the expression of ATM, JAK and STAT3, respectively. RT-qPCR, western blot and cell migration assay were performed at 6, 12 and 24 h respectively.

Zhang L., Xu L.J., Zhu J., Li J., Xue B.X., Gao J., Sun C.Y., Zang Y.C., Zhou Y.B., Yang D.R, & Shan Y.X. (2018). ATM-JAK-PD-L1 signaling pathway inhibition decreases EMT and metastasis of androgen-independent prostate cancer. Molecular Medicine Reports, 17(5), 7045-7054.

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Tumor Cell Invasion Assay with Inhibitors

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Tumor cells (1×10⁴) were seeded into the upper chamber of a Transwell support (Corning Incorporated, Corning, NY, USA) with serum-free media, whereas medium containing 10% FBS was added to the lower chamber. Following incubation for 24 h at 37°C, cells were fixed in methanol and then stained with crystal violet. Positively stained cells in three randomly selected visual fields were counted under a light microscope (Olympus Corporation), magnification, ×20 (inlet, ×100). Each assay was repeated three times. Antibodies used were: JAK inhibitor 1 (420099; EMD Millipore), CP466722 (S2245; Selleck Chemicals), PD-L1 Antibody (62-5982-80, Thermo Fisher Scientific, Inc.).

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