The following antibodies were purchased from Abcam: OLA1 (ab229090), GAPDH (ab8245), CCNB1 (ab71977), CCNB2 (ab18250), CCNE2 (ab40890), CDK2 (ab32147), CDK4 (ab108357), P21 (ab188224), Bcl-2 (ab32124), BAD (ab32445), cleaved caspase 3 (ab32042), Rb (ab181616), pRb (ab184796), and E2F1 (ab112580). The P21 inhibitor UC2288 was purchased from Abcam (ab146969) and used as recommended.
Uc2288
Manufactured by Abcam
Sourced in United Kingdom
UC2288 is a lab equipment product that serves as a core function. It is designed for use in laboratory settings, but a detailed description of its intended use is not available.
Automatically generated - may contain errors
Lab products found in correlation
Ab32147, by Abcam (1 mentions)
Ab40890, by Abcam (1 mentions)
Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (1 mentions)
Epidermal growth factor (egf), by Abcam (1 mentions)
N2 supplement, by Thermo Fisher Scientific (1 mentions)
Doxorubicin, by Merck Group (1 mentions)
Torin, by Bio-Techne (1 mentions)
Arpe 19, by American Type Culture Collection (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
Cm h2dcfda, by Thermo Fisher Scientific (1 mentions)
L ng nitroarginine methyl ester l name, by Merck Group (1 mentions)
L arginine l arg, by Merck Group (1 mentions)
N acetylcysteine, by Merck Group (1 mentions)
Ascorbic acid, by Merck Group (1 mentions)
Potassium iodide, by Merck Group (1 mentions)
Phosphoric acid, by Merck Group (1 mentions)
Nitric oxide assay kit, by Abcam (1 mentions)
Goat anti rabbit igg hrp, by Cell Signaling Technology (1 mentions)
P erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions)
Survivin, by Cell Signaling Technology (1 mentions)
5 protocols using uc2288
1
Protein Expression and Apoptosis Pathway Analysis
2
Culturing and Differentiating Brain Cells
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Brain cells were grown in Gibco Dulbecco’s modified Eagle’s medium (DMEM)/F-12 medium (Thermo Fisher Scientific, Waltham, MA, USA) containing basic fibroblast growth factor (bFGF; 20 ng/mL), epidermal growth factor (EGF; 20 ng/mL; PeproTech, Rocky Hill, NJ, USA), 1% N-2 supplement (Thermo Fisher Scientific, Waltham, MA, USA), penicillin–streptomycin, and 2% fetal bovine serum for 1 week and moved into the low attachment dish (Alpha Plus Scientific, Taoyuan, Taiwan) with floating cultures at the density of 7–8 cells/µL in DMEM/F-12, EGF, fibroblast growth factor 2, and N-2 supplement for 21 days.25 (link) To treat the primary pericytes with p21 inhibitor, pericytes were incubated with 10 µM UC2288 (Abcam, Cambridge, UK) for 24 h before being grown in a suspension culture system. For a single experiment, at least four animals were required for the sphere forming assay.
3
Cellular Response to Doxorubicin and mTOR Inhibitors
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HeLa and ARPE-19 cells were purchased from ATCC. ARPE-19 is a spontaneously immortalized cell line of human retinal pigment epithelium. These cells are diploid and non-transformed. HeLa TFEB and TFEB/TFE3 knockout and p53-null human osteosarcoma (SAOS-2 p53-null) cells were kindly provided by Richard J. Youle (National Institutes of Health, Bethesda, MD, USA) and Dr. Caterina Missero (Centro di Ingegneria Genetica e Biotecnologie Avanzate, Naples, Italy), respectively. HeLa cells stably expressing TFEB-GFP or TFEB 3xflag were obtained as previously described [22 (link),28 (link)]. Cells were cultured at 37 °C and 5% CO2 in DMEM (Sigma-Aldrich, St. Louis, MO, USA, D6429) supplemented with fetal bovine serum (10%) and Pen/Strep (1%). Treatments with doxorubicin (D1515, Sigma, St. Louis, MO, USA, D6429), Torin (Tocris, Bristol, UK), or UC2288 (ab146969, Abcam, Cambridge, UK) were performed at indicated time points and concentrations and compared with control DMSO. For colony formation assay, about 200 cells per well were seeded in a 6-well plate. Next day cells were treated, and maintained in culture 8 days more, replacing medium every three days.
4
Antioxidant and Nitric Oxide Modulation
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N-acetylcysteine (NAC), L-NG-nitroarginine methyl ester (L-NAME), iodine, potassium iodide, phosphoric acid, ascorbic acid, L-arginine (L-Arg), and BH4 were purchased from Sigma-Aldrich (St. Louis, MO, USA). CM-H2DCFDA was obtained from Thermo Fisher Scientific (Waltham, MA, USA). The Nitric oxide assay kit and UC2288 were purchased from Abcam (Cambridge, UK).
5
Investigating EGFR Signaling Pathways
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UC2288 (purity ~99%) and human recombinant epidermal growth factor (EGF) were purchased from Abcam (Cambridge, UK), and dissolved in dimethyl sulfoxide (DMSO) and water, respectively. Cisplatin (Haosen, Jiangsu, China) was dissolved in water. The primary antibodies including EGFR, p-EGFR (Tyr1068), ERK1/2, p-ERK1/2 (Thr202/Tyr204), caspase 3, cleaved-caspase 3, γ-H2AX, survivin, Bax, Bcl-2, GAPDH and goat anti-rabbit IgG-HRP and Alexa Fluor-488-labeled goat anti-rabbit (IgG) secondary antibody secondary antibodies were obtained from Cell Signaling Technology (Massachusetts, USA).
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