U0126 9903

Manufactured by Cell Signaling Technology

Sourced in United States, Germany

U0126 (9903) is a small molecule inhibitor that selectively inhibits the activation of MEK1 and MEK2, key enzymes in the ERK signaling pathway. It is commonly used as a research tool to study the role of the ERK pathway in various cellular processes.

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Lab products found in correlation

Protein g agarose, by Roche (1 mentions) Block it pol 2 mir rnai expression vector kit, by Thermo Fisher Scientific (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Glutathione sepharose 4b, by GE Healthcare (1 mentions) Pf573228 sc 204179, by Santa Cruz Biotechnology (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) All in one qpcr mix, by GeneCopoeia (1 mentions) All in one first strand cdna synthesis kit, by GeneCopoeia (1 mentions) Sb203580 5633, by Cell Signaling Technology (1 mentions) Hrp conjugated anti rabbit igg 7074, by Cell Signaling Technology (1 mentions) Mouse anti gapdh nb300 221 antibody, by Novus Biologicals (1 mentions) Recombinant human tnfα 210 ta, by R&D Systems (1 mentions) Gapdh 2118, by Cell Signaling Technology (1 mentions) Temozolomide, by Merck Group (1 mentions) Pimozide, by Merck Group (1 mentions) Saracatinib, by Selleck Chemicals (1 mentions) Recombinant human egf, by Thermo Fisher Scientific (1 mentions) Gefitinib s1025, by Selleck Chemicals (1 mentions) γ 32p atp, by PerkinElmer (1 mentions) 3h leucine, by PerkinElmer (1 mentions)

Spelling variants of this product

U0126 (515 citations)

6 protocols using u0126 9903

Antibody Immunoprecipitation and Western Blot Analysis

Cited 1 time

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Protein-G agarose (11243233001) was obtained from Roche. Glutathione Sepharose 4B (17-0757-01) was from GE Healthcare. Goat anti-mouse IgG conjugated with Alexa Fluor 488 (A21026), goat anti-rabbit IgG conjugated Alexa Fluor 594 (A11012), and blasticidine (R210-01) were from Invitrogen. Recombinant human FAK protein (31168) was from Active Motif (Carlsbad, CA). PF573228 (sc-204179) was from Santa Cruz. U0126 (9903) was from Cell Signaling. Protease inhibitor cocktail (P8340), 17-AAG (A8476), Lubrol-PX, DMSO, and other chemicals were from Sigma (St. Louis, MO). HSP90β miRNAi constructs were generated using the BLOCK-iT Pol II miR RNAi Expression Vector Kit (Invitrogene, K4936-00). Oligonucleotide sequences for miRNAi constructs are 90β-1205 5′-ATA AAG TTG AGG TAC TCA GGT-3′ (31 (link)), and FAK-1763 5′-ATC ATT TGA AGA CAC CAG AAC-3′ (32 (link)). A scrambled sequence shRNA that is not homologous to any human gene was used as control. Antibodies were purchased from Santa Cruz (FAK, sc-558; actin, sc-1616), Cell Signaling (p-Tyr397-FAK, 3283S; p-Tyr118-paxillin, 2541; B-Raf, 9434; Raf-1, 9422), Invitrogen (HSP90β, 37-9400; Myc, 132500), Chemicon international (GAPDH, MAB374), Thermo Scientific (rabbit IgG, 31460; mouse IgG, 31430), Sigma (Flag, F3165; paxillin, P1039).

Xiong X., Wang Y., Liu C., Lu Q., Liu T., Chen G., Rao H, & Luo S. (2014). Heat Shock Protein 90β Stabilizes Focal Adhesion Kinase and Enhances Cell Migration and Invasion in Breast Cancer Cells. Experimental cell research, 326(1), 78-89.

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Molecular Assays for Cell Signaling

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α-Cyano-4-hydroxycinnamic acid (α-CHCA) (C2020) was obtained from Sigma (St. Louis, MO, USA). BAY 11-7085 (HY-10257) was purchased from MedChemExpress, and U0126 (9903) was from Cell Signaling Technology (Danvers, MA, USA). Medium and fetal bovine serum (FBS) were purchased from Thermo Scientific (Kalamazoo, MI, USA). LDH (ab102526) was purchased from Abcam. Trizol was from Invitrogen (Invitrogen, Thermo Fisher Scientific). Antibodies were obtained from Abclonal. All-in-one™ first-strand cDNA synthesis kit and All-in-one™ qPCR mix were from Genecopoeia™ (Rockville, MD, USA). Other chemical reagents were from Sigma (St. Louis, MO, USA).

Wang Y., Zhou X., Zou K., Chen G., Huang L., Yang F., Pan W., Xu H., Xu Z., Chen H., Chen J., Gong S., Zhou X., Xu W, & Zhao J. (2021). Monocarboxylate Transporter 4 Triggered Cell Pyroptosis to Aggravate Intestinal Inflammation in Inflammatory Bowel Disease. Frontiers in Immunology, 12, 644862.

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Signaling Pathways Activation in TNF-α and CCL21 Response

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Recombinant human TNF‐α (210‐TA) and CCL21 (366‐6C) were purchased from R&D Systems (Minneapolis, MN, USA). The following antibodies were used in Western blot analyses and immunocytochemistry: rabbit anti‐TNF‐α (ab6671), goat anti‐rabbit FITC‐conjugated IgG H&L (ab6717), and rabbit anti‐CCR7 (ab103404) from Abcam (Cambridge, MA, USA); rabbit anti‐p38 (8690), rabbit anti‐phospho‐p38 (Thr180/Tyr182; 4511), rabbit anti‐ERK (9102), rabbit anti‐phospho‐ERK (Thr202/Tyr204; 9101), and HRP‐conjugated anti‐rabbit IgG (7074) antibodies from Cell Signaling Technology (Danvers, MA, USA); and mouse anti‐GAPDH (NB300‐221) antibody from Novus Biologicals (Littleton, CO, USA). The ERK inhibitor U0126 (9903) and the p38 inhibitor SB203580 (5633) were purchased from Cell Signaling Technology.

Maolake A., Izumi K., Natsagdorj A., Iwamoto H., Kadomoto S., Makino T., Naito R., Shigehara K., Kadono Y., Hiratsuka K., Wufuer G., Nastiuk K.L, & Mizokami A. (2018). Tumor necrosis factor‐α induces prostate cancer cell migration in lymphatic metastasis through CCR7 upregulation. Cancer Science, 109(5), 1524-1531.

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Analyzing EGFR and Downstream Signaling

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Phospho-EGFR (3777, 2231), EGFR (4267), phospho-Src (6943), Src (2109), phospho-p44/42 (4370), p44/42 (9102), phospho-Stat3 (9145), Stat3 (4904), phospho-Stat5 (4322), Stat5 (9363), Fn14 (4403), Cleaved Caspase 3 (9661), γH2AX (9718), HA (2367), and GAPDH (2118) were from Cell Signaling Technology. Antibodies to α-tubulin and β-actin were from Millipore.
Human recombinant EGF was purchased from PeproTech. Temozolomide and Pimozide (P1793) were obtained from Sigma. U0126 (9903) was purchased from Cell Signaling Technology. Erlotinib (S7786), Gefitinib (S1025), and Saracatinib (S1006) were purchased from Selleck Chem.

Roos A., Dhruv H.D., Peng S., Inge L.J., Tuncali S., Pineda M., Millard N., Mayo Z., Eschbacher J.M., Loftus J.C., Winkles J.A, & Tran N.L. (2018). EGFRvIII-Stat5 Signaling Enhances Glioblastoma Cell Migration and Survival. Molecular cancer research : MCR, 16(7), 1185-1195.

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Quantifying HNE-Induced Protein Synthesis

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4-hydroxynonenal (HNE) was obtained from EMD Millipore (Billerica, MA, USA). The MEK1/2 inhibitor U0126 (9903) was purchased from Cell Signaling Technology and the mTORC1 inhibitor Rapamycin (R-5000) was obtained from LC Laboratories (Woburn, MA). The [³H]-leucine and the [γ-³²P]-ATP were purchased from Perkin Elmer (Boston, MA). All other materials were purchased from Thermo Fisher Scientific (Waltham, MA, USA) unless otherwise specified.

Calamaras T.D., Lee C., Lan F., Ido Y., Siwik D.A, & Colucci W.S. (2015). Lipid peroxidation product 4-Hydroxy-trans-2-nonenal (HNE) causes protein synthesis in cardiac myocytes via activated mTORC1-P70S6K-RPS6 signaling. Free radical biology & medicine, 82, 137-146.

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Lipid Signaling Pathway Inhibitor Assay

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LPA 16:0 (857123P), 18:0 (857128P) and 18:1 (857130P) was obtained from Avanti Polar Lipids (Alabaster, AL, USA), LPA 18:2 (L-0182) and 20:4 (L-0204) from Echelon Bioscience (Salt Lake City, UT, USA). Ro6842262 (5913), H2L5186303 (4878) were purchased from Tocris Biosciences (Bristol, UK); Y27632 (10005583), HA1077 (10010559), RKI1447 (16278), Gö6983 (13311), BIRB796 (Doramapimod, 104640), from Cayman Chemicals (Ann Arbor, MI, USA); CRT0066101 (SML1507) from Sigma-Aldrich (Taufkirchen, Germany); U0126 (9903) from Cell Signaling (Frankfurt, Germany). All inhibitors, their targets and the concentrations used are listed in Table 1.

Ojasalu K., Lieber S., Sokol A.M., Nist A., Stiewe T., Bullwinkel I., Finkernagel F., Reinartz S., Müller-Brüsselbach S., Grosse R., Graumann J, & Müller R. (2023). The lysophosphatidic acid-regulated signal transduction network in ovarian cancer cells and its role in actomyosin dynamics, cell migration and entosis. Theranostics, 13(6), 1921-1948.

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