U0126

Manufactured by Santa Cruz Biotechnology

Sourced in United States

U0126 is a selective inhibitor of the extracellular signal-regulated kinases 1 and 2 (ERK1/2). It functions by blocking the activation of MEK1/2, the kinases responsible for phosphorylating and activating ERK1/2.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

ERK1/2 inhibitor (U0126) (3 citations) U0126 (sc-222395) (2 citations)

33 protocols using u0126

Antibody and Drug Preparation for Cell Studies

Check if the same lab product or an alternative is used in the 5 most similar protocols

A rabbit anti-human Kir2.1 polyclonal antibody was purchased from Alomone labs (Jerusalem, Israel), andpolyclonal anti-MRP1/ABCC1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibodies were purchased from Santa Cruz Inc. (CA, USA). Horseradish peroxidase (HRP)-labeled goat anti-rabbit immunoglobulin G (IgG) and goat anti-mouse IgG were obtained from Santa Cruz Inc. U0126, the MEK inhibitor, and staurosporine, the PKC inhibitor, were purchased from Selleck Chemicals (Houston, TX, USA). All three chemotherapeutic drugs, Cisplatin (DDP; Shandong, China), Etoposide (VP-16; Jiangsu, China) and Adriamycin (ADM; Jiangsu, China), were obtained from commercial sources and were dissolved according to the manufacturer’s instructions.

Liu H., Huang J., Peng J., Wu X., Zhang Y., Zhu W, & Guo L. (2015). Upregulation of the inwardly rectifying potassium channel Kir2.1 (KCNJ2) modulates multidrug resistance of small-cell lung cancer under the regulation of miR-7 and the Ras/MAPK pathway. Molecular Cancer, 14, 59.

+ Open protocol

+ Expand

Glioma Cell Line Characterization

Check if the same lab product or an alternative is used in the 5 most similar protocols

The human glioma U-251 cell line was kindly provided by Dr. Dah-Yu Lu (Department of Pharmacology, School of Medicine, China Medical University, Taichung, Taiwan). The human glioma GBM8401 and GBM8901 cell line was kindly provided by Dr. Li-Sung Hsu (Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan). The U-87MG (BCRC No. 60360) and M059K cell line (BCRC No. 60381) were purchased from the Bioresources Collection and Research Center (BCRC, Hsinchu, Taiwan). These cells were cultured in DMEM/F12 and MEM medium supplemented with 1 × penicillin/streptomycin and 10% fetal bovine serum (HyClone, Logan, UT, USA) were incubated at 37 °C in a humidified incubator containing a 5% CO₂ atmosphere. LCN2 antibody was purchased from R&D Systems, Inc (Minneapolis, MN, USA). U0126 (MEK inhibitor), siRNA-CTSD (si-CTSD), Cathepsin D (CTSD), t-ERK, β-actin, HRP-mouse and HRP-rabbit antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Matrigel Matrix was purchased from Corning company (Tewksbury, MA, USA). Phosphorylated-ERK (p-ERK) was obtained from Cell Signaling Technology (Beverly, MA, USA).

Hsieh Y.H., Tsai J.P., Yu C.L., Lee C.C., Hsu J.C, & Chen J.C. (2021). Overexpression of Lipocalin-2 Inhibits Proliferation and Invasiveness of Human Glioblastoma Multiforme Cells by Activating ERK Targeting Cathepsin D Expression. Biology, 10(5), 390.

+ Open protocol

+ Expand

Anti-KIR2DL4 Antibody Signaling Pathway

Check if the same lab product or an alternative is used in the 5 most similar protocols

The anti-KIR2DL4 agonistic antibody (mouse monoclonal IgG, Clone; 181703) was purchased from R&D Systems (Minneapolis, MN). Another anti-KIR2DL4 antibody (rabbit IgG, polyclonal, ab154386) and an anti-phospho-SHP-2 antibody (Y542, rabbit polyclonal IgG), used for immunostaining and Western blotting were obtained from Abcam (Cambridge, MA). An anti-phospho-ERK antibody and an anti-GAPDH antibody were obtained from Cell Signaling Technology (Beverly, MA). The KIR2DL4-targeted shRNA lentiviral particles and the control particles were purchased from Santa Cruz Biotechnology (San Diego, CA). Infection of these viral particles and the following selection were performed according to the manufacturer's instructions. The specific MAP2K1 inhibitor U0126, the specific ERK inhibitor FR180204, and the specific SHP-2 inhibitor PHPS1 were purchased from Santa Cruz Biotechnology [22 (link)–24 (link)]. We used all inhibitors at concentrations of 10 µM.

Takei Y., Ueshima C., Kataoka T.R., Hirata M., Sugimoto A., Rokutan-Kurata M., Moriyoshi K., Ono K., Murakami I., Iwamoto S, & Haga H. (2017). Killer cell immunoglobulin-like receptor 2DL4 is expressed in and suppresses the cell growth of Langerhans cell histiocytosis. Oncotarget, 8(23), 36964-36972.

+ Open protocol

+ Expand

Isolation and Culture of Mouse Hepatocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Hepatocytes were isolated from wild-type, 6–12 week-old C57BL/6 mice (Jackson Laboratory, Bar Harbor, ME). In some experiments, hepatocytes were isolated from Hnf4a^fl/fl:Alb Cre⁺ mice that lacked HNF4α expression in adult hepatocytes and Hnf4a^fl/fl:Alb Cre⁻ wild-type littermates (kind gift from Dr. Frank J. Gonzalez) (17 (link)). All mice were cared for in accordance to the National Institutes of Health “Guide for the Care and Use of Laboratory Animals.” Hepatocyte isolation was performed by two-step perfusion using Liver Perfusion and Liver Digest Media (Life Technologies, Pleasanton, CA) followed by separation with 50% Percoll (GE Healthcare Life Sciences, Pittsburgh, PA) density gradient. Purity of live hepatocytes was routinely ≥90% by trypan blue exclusion. Hepatocytes were cultured in 5% fetal calf serum (Hyclone, Logan, UT) in DMEM supplemented with L-glutamine, antibiotics, insulin-transferrin-selenium, and HEPES (Mediatech, Manassas, VA). Inhibitors used and their final concentrations in cell culture were: 50μM Y-27632, 25μM FAK inhibitor-14, 0.5μM blebbistatin, or 20μM U-0126 (Santa Cruz Biotech, Dallas, TX). Unless otherwise noted, functional assays and gene expression analysis were performed 24h after hepatocyte plating.

Desai S.S., Tung J.C., Zhou V.X., Grenert J.P., Malato Y., Rezvani M., Español-Suñer R., Willenbring H., Weaver V.M, & Chang T.T. (2016). Physiological Ranges of Matrix Rigidity Modulate Primary Mouse Hepatocyte Function In Part Through Hepatocyte Nuclear Factor 4 Alpha. Hepatology (Baltimore, Md.), 64(1), 261-275.

+ Open protocol

+ Expand

Investigating MAPK Signaling in Breast Cancer

Check if the same lab product or an alternative is used in the 5 most similar protocols

Recombinant human IL1B (200-01B) and IL-1R antagonist (IL1-RA, 200-01R) were purchased from Peprotech (Rocky Hill, NJ). p38 MAPK (8690), phospho-p38 MAPK (Thr180/Tyr182; 4511), p42/44 MAPK (9107S), phospho-p42/44 MAPK (Thr202/Tyr204; 9101) antibodies were purchased from Cell Signaling Technology (Beverly, MA). MAPK inhibitors SP600125, SB202190 and SB203580 were purchased from Sigma Aldrich (St Louis, MO), U0126 and BAY869766 were purchased from Santa Cruz Biotechnologies (Santa Cruz, CA).
The human breast cancer lines: MDA-MB-231, MDA-MB-436, BT549, SKBR3, ZR75-1, HCC1954 were cultured in Dulbecco’s Modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals, Lawrenceville, GA), 2 mM L-glutamine, and 50 μg/mL gentamicin (Life Technologies, Carlsbad, CA). THP-1 monocyte cells were cultured in RPMI 1640 supplemented with 10% FBS, 2 mM L-glutamine, 1 mM sodium pyruvate, 10 mM HEPES, and 1% antibiotic/antimycotic solution (15240062, Life Technologies). All cell lines were recently acquired from the ATCC (Manassas, VA). Cell lines were incubated in a humidified atmosphere of 5% CO₂ at 37 °C.

Chung S.T., Geerts D., Roseman K., Renaud A, & Connelly L. (2017). Osteoprotegerin mediates tumor-promoting effects of Interleukin-1beta in breast cancer cells. Molecular Cancer, 16, 27.

+ Open protocol

+ Expand

Molecular Pathways in Fibrosis Regulation

Check if the same lab product or an alternative is used in the 5 most similar protocols

JQ1 was purchased from Tocris Bioscience (Bristol, UK). The recombinant human TGF-β1 was from PeproTech (Rocky Hill, NJ, USA). Antibodies used in Western blot and sources were as follows. The rabbit anti-Brd4 antibody was supplied by Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit anti-α-SMA, anti-fibronectin and anti-collagen IV antibodies were purchased from Abcam (Cambridge, UK). Rabbit anti-Smad3, anti-p-Smad3, anti-ERK1/2, anti-p-ERK1/2 and anti-Nox4 antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). The rabbit anti-GAPDH antibody was from Bioss (Beijing, China). DCFH-DA was from Beyotime Biotechnology (Jiangsu, China), Amplex Red Hydrogen Peroxide/Peroxidase Assay Kit from Molecular Probes (Eugene, OR, USA), polyethylene glycol (PEG)–catalase and N-acetyl-cysteine (NAC) from Sigma-Aldrich (St. Louis, MO, USA), SIS3 from Cayman Chemical (Ann Arbor, MI, USA), and U0126 from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Zhou B., Mu J., Gong Y., Lu C., Zhao Y., He T, & Qin Z. (2016). Brd4 inhibition attenuates unilateral ureteral obstruction-induced fibrosis by blocking TGF-β-mediated Nox4 expression. Redox Biology, 11, 390-402.

+ Open protocol

+ Expand

Resveratrol Modulates Egr-1 Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

The final concentrations of all the inhibitors were 10 μM except SP 600125 (BIOMOL International, PA), of which the concentration was 20 μM. SB 203580 (Promega, WI) inhibits p38MAPK/SAP2, whereas SP 600125 is a potent JNK inhibitor. U-0126 (Santa Cruz Biotechnology, Inc, TX) is a potent inhibitor of ERK1/2 activity and LY 294002 (Santa Cruz Biotechnology, Inc, TX) blocks PI3K/AKT pathways. Cells were treated with or without 100 μM of resveratrol for another 6 h after 1 h preincubation, and the expression of Egr-1 protein and β-actin (loading control) was examined by western blotting.

Shi Q., Geldenhuys W., Sutariya V., Bishayee A., Patel I, & Bhatia D. (2015). CArG-driven GADD45α activated by resveratrol inhibits lung cancer cells. Genes & Cancer, 6(5-6), 220-230.

+ Open protocol

+ Expand

CRISPR-Mediated ATF3 Knockout in BEAS-2B Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human bronchial epithelial BEAS-2B cells were purchased from the China Center for Type Culture Collection (CCTCC, Wuhan, China) and cultured in Dulbecco’s Modified Eagle Medium (DMEM, Invitrogen, San Diego, CA, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, NY, USA). ATF3 KO BEAS-2B cells were custom developed by using a CRISPR/Cas9 gene-editing method by Cyagen Biosciences (Suzhou, China). Sodium meta-arsenite (NaAsO₂) was purchased from Sigma (St Louis, MO, USA). The inhibitors, SB203580, SP600125, U0126 and LY294002, were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Antibodies against ATF3 (33593), DR5 (3696), JNK (9252), p-JNK (4668), p38 (8690), p-p38 (9215), Bcl-xL (2764), caspase 8 (9746), cleaved caspase 8 (8592), caspase 3 (14220), cleaved caspase 3 (9664), caspase 9 (9502), cleaved caspase 9 (9509), α-tubulin (2125), AKT (C67E7), p-AKT (4060T), ERK (4695), p-ERK (91015) and β-actin (4970) were purchased from Cell Signaling Technologies (Danvers, MA, USA).

Shi Q., Hu B., Yang C., Zhao L., Wu J, & Qi N. (2021). ATF3 Promotes Arsenic-Induced Apoptosis and Oppositely Regulates DR5 and Bcl-xL Expression in Human Bronchial Epithelial Cells. International Journal of Molecular Sciences, 22(8), 4223.

+ Open protocol

+ Expand

Muscle Stem Cell Differentiation Modulation

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were plated in muscle growth media (fetal bovine serum 0.05 mL/mL, fetuin 50 μg/mL, epidermal growth factor 10 ng/mL, basic fibroblast growth factor 1 ng/mL, insulin 10 μg/mL, and dexamethasone 0.4 μg/mL) for 48 hours before changing to skeletal muscle differentiation media, which is a proprietary serum-free medium containing 10 μg/mL insulin (PromoCell) for 14 days. PMSCs were treated every 3 days with 200 nM of IGF-IR inhibitor PPP, 25 μM of AKT inhibitor LY294002, 10 μM of MEK1/2 inhibitor U0126, or 10 μM of IR inhibitor HNMPA (Santa Cruz Biotechnology) under muscle differentiation conditions. Treatment concentrations for LY294002, U0126, and HNMPA were determined by a dose-response experiment using PMSCs in muscle differentiation media (Supplementary Figure 1). For IGFBP-6 supplementation with the inhibitors, recombinant human IGFBP-6 (ProSpec) was added to the media (375 ng/mL) every 3 days at the time of media change. The dose of IGFBP-6 was based on our previous studies [26 (link), 31 (link)].

Aboalola D, & Han V.K. (2019). Insulin-Like Growth Factor Binding Protein-6 Promotes the Differentiation of Placental Mesenchymal Stem Cells into Skeletal Muscle Independent of Insulin-Like Growth Factor Receptor-1 and Insulin Receptor. Stem Cells International, 2019, 9245938.

+ Open protocol

+ Expand

Quantifying Protein Signaling Cascades

Check if the same lab product or an alternative is used in the 5 most similar protocols

TNF-α was obtained from ProSpec-Tany TechnoGene Ltd. (cyt-223-b) and U0126 was purchased from Santa Cruz (CAS 109511–58-2). The antibodies used were: rabbit anti-phospho-ERK1/2 (T202/Y204, Cell Signaling), mouse anti-ERK1/2 (L34F12, Cell Signaling), rabbit anti-MMP-9 (AB19016, Merck Millipore), rabbit anti-MMP2 (VARP20016_T100, Aviva), rabbit anti-MMP14 (PA5–13,183, Thermo Scientific), rabbit anti-FAP-α (GTX102732, GeneTex), goat anti-DPPIV/CD26 (AF1180, R&D Systems), rabbit-anti-Cortactin (H-191) (sc-11,408, Santa Cruz Biotechnology, Inc.), goat anti-actin (I-19) (sc-1616, Santa Cruz Biotechnology, Inc.) and mouse anti-FL-2 (B6) (sc-28,320, Santa Cruz Biotechnology, Inc.). The cholera toxin subunit B-HRP conjugate was obtained from Molecular Probes (C-34,780). Millicell Hanging Cell Culture Inserts (8.0 μm) were purchased from Merck Millipore (PIEP15R48) and blotting membranes were purchased from Bio-Rad (162–0094). Immuno-detection was performed using HRP-conjugated donkey anti-rabbit (sc-2313), donkey anti-goat (sc-2020) and goat anti-mouse (sc-2005) antibodies purchased from Santa Cruz Biotechnology, Inc. and Pierce™ECL Western Blotting Substrate (32,106, Thermo Scientific).

Wolczyk D., Zaremba-Czogalla M., Hryniewicz-Jankowska A., Tabola R., Grabowski K., Sikorski A.F, & Augoff K. (2016). TNF-α promotes breast cancer cell migration and enhances the concentration of membrane-associated proteases in lipid rafts. Cellular Oncology (Dordrecht), 39, 353-363.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!