Log In Sign up
The largest database of trusted experimental protocols

Mek inhibitor u0126

Manufactured by Promega
Visit Supplier
Sourced in United States

U0126 is a selective and potent inhibitor of the MAP kinase kinases MEK1 and MEK2. It blocks the activation of ERK1 and ERK2 by inhibiting the enzymes that phosphorylate and activate them.

Automatically generated - may contain errors

Lab products found in correlation

Erdafitinib, by Selleck Chemicals (1 mentions) Byl719, by Selleck Chemicals (1 mentions) Signalsilence p44 42 mapk erk1 2 sirna, by Cell Signaling Technology (1 mentions) Mopc 21, by BioLegend (1 mentions) Perm wash, by BD (1 mentions) Pd173074, by Selleck Chemicals (1 mentions) Cytofix cytoperm, by BD (1 mentions) Cytoflex lx flow cytometer, by Beckman Coulter (1 mentions) Cytexpert, by Beckman Coulter (1 mentions) Cell lysis buffer, by Cell Signaling Technology (1 mentions) Pathscan phospho s6 ribosomal protein ser235 236 sandwich elisa kit, by Cell Signaling Technology (1 mentions) Infinite m200 plate reader, by Tecan (1 mentions) Hydrogen peroxide, by Thermo Fisher Scientific (1 mentions) Spectramax m3 multi mode microplate reader, by Molecular Devices (1 mentions) Sb203580, by Promega (1 mentions) Ly294002, by Promega (1 mentions) Pd98059, by Promega (1 mentions) Sch772984, by Adooq (1 mentions) 5 aza 2 deoxycytidine, by Merck Group (1 mentions) Gapdh antibody, by Bioworld Technology (1 mentions)

5 protocols using mek inhibitor u0126

1

Immune Cell Profiling and Modulation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Immune cells from mice were stained with PerCP-conjugated anti-CD4 (GK1.5, BioLegend) mAbs, APC/Cy7-conjugated anti-CD8a (53–6.7) mAbs, and the isotype monoclonal mAb. FITC-conjugated anti-I-A/I-E mAbs (M5/114.15.2, BioLegend) was used for negative gating. After pretreatment with 3 μM FGFR1-TKIs (PD173074; AZD4547; Erdafitinib, Selleck Chemicals), 3 μM mitogen-activated protein kinase (MAPK) inhibitor (MEK inhibitor U0126, Promega), MAPK siRNA (SignalSilence® p44/42 MAPK Erk1/2 siRNA, Cell Signaling Technology), 3 μM STAT3 inhibitor (S3I-201, Selleck Chemicals), or 3 µM PI3K inhibitor (BYL719, Selleck Chemicals) for 48 hr, HLA class I and HLA-DR expression on tumor cell lines was assessed via flow cytometry using anti-HLA class I antibodies (Abs) conjugated with fluorescein isothiocyanate (G46-2, BD Pharmingen) and anti-HLA-DR Abs conjugated with phycoerythrin (TU36, BD Pharmingen). HNSCC cell lines were treated with or without 50 U/ml IFN-γ for 48 hr before the assay. IgG1 (MOPC-21, BioLegend) and IgG2a (MOPC-173; BioLegend) were used as isotype controls. Intracellular IFN-γ staining were performed using Perm/WashTM (BD Pharmingen), Cytofix/CytopermTM (BD Pharmingen), APC-conjugated anti-IFN-γ mAbs (4S.B3, BioLegend), and FITC-conjugated anti-granzyme B mAbs (GB11, BioLegend). Samples were analyzed using the CytoFLEX LX flow cytometer and CytExpert (Beckman Coulter).
Kono M., Komatsuda H., Yamaki H., Kumai T., Hayashi R., Wakisaka R., Nagato T., Ohkuri T., Kosaka A., Ohara K., Kishibe K., Takahara M., Katada A., Hayashi T., Kobayashi H, & Harabuchi Y. (2022). Immunomodulation via FGFR inhibition augments FGFR1 targeting T-cell based antitumor immunotherapy for head and neck squamous cell carcinoma. Oncoimmunology, 11(1), 2021619.
+ Open protocol
+ Expand
2

Profiling mTOR Pathway Activation in MEF Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
MEF cell cultures were incubated in serum-free medium for 24 h before the experiment. Then cells were washed with cold PBS, and lysed with cell lysis buffer from Cell Signaling Technology (#9803), scraped and sonicated briefly. Then the extracts were centrifuged for 10 min at 14,000 × g at 4 °C. The supernatants were aliquoted and stored at −80 °C until used. For the experiments with inhibitors, cells were treated 30 min before harvest with 10 μM MEK inhibitor U0126 (Promega), 50 μM PI3 kinase inhibitor LY294002 (Cell Signaling Technology) and 20 nM mTOR inhibitor rapamycin (Calbiochem). Phosphorylation assays were performed by using PathScan®Phospho-S6 Ribosomal Protein (Ser235/236) Sandwich ELISA kit (#7205) and PathScan®Phospho-4E-BP1 (Thr37/Thr46) Sandwich ELISA kit (#7216) (Cell Signaling Technology) from the aliquots collected. For the analysis of Total-S6 or Total-4E-BP1 protein levels, PathScan®Total-S6 Ribosomal Protein Sandwich ELISA kit (#7225) and PathScan®Total-4E-BP1 Sandwich ELISA kit (#7179) were used (Cell Signaling Technology). The analyses of the ELISAs were performed on the Tecan Infinite M200 plate reader.
Lastres-Becker I., Nonis D., Eich F., Klinkenberg M., Gorospe M., Kötter P., Klein F.A., Kedersha N, & Auburger G. (2016). MAMMALIAN ATAXIN-2 MODULATES TRANSLATION CONTROL AT THE PRE-INITITATION COMPLEX VIA PI3K/MTOR AND IS INDUCED BY STARVATION. Biochimica et biophysica acta, 1862(9), 1558-1569.
+ Open protocol
+ Expand
3

Estrogen-Induced ROS Production Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were plated at 10,000 cells/well in a 96-well plate, allowed to attach overnight, and then treated with 100 μl of media containing 1% four times charcoal-stripped FBS for 48 hr. Cells were loaded with 15μM 2’,7’-Dichlorodihydrofluorescein diacetate (DCDHF) (Enzo Life Sciences) for 1 hr. Then the production of ROS was measured in cells after E2 or DES treatment for 15 min. Hydrogen peroxide (1μM, Fisher Scientific – Pittsburg, PA) and ethanol (0.0001%) were used as positive and negative controls, respectively. For both E2 and DES treatments, the concentrations spanned 10−14 to 10−6 M. Dichlorofluorescein production, formed as a result of ROS/DCDHF interaction, was measured at an excitation of 485 nm, and an emission of 538 nm in a SpectraMax M3 Multi-Mode Microplate Reader (Molecular Devices). For studies with MEK inhibitor U0126 (Promega – Madison, WI), cells were co-incubated with 10−7M inhibitor during the last 30 min of the DCDHF incubation.
Koong L.Y, & Watson C.S. (2014). Direct estradiol and diethylstilbestrol actions on early- vs. late-stage prostate cancer cells. The Prostate, 74(16), 1589-1603.
+ Open protocol
+ Expand
4

Signaling Pathway and Epigenetic Modulation Experiments

Check if the same lab product or an alternative is used in the 5 most similar protocols
For signaling pathway inhibition experiments, cells were treated with vehicle (0.1% DMSO) or PI3K inhibitor LY294002 (50 μm) (Promega, Madison, WI, USA), p38 MAPK inhibitor SB203580 (10 μm) (Promega), MEK inhibitor U0126 (10 μm) (Promega), PD98059 (20 μm) (Promega), Raf‐1 kinase inhibitor ZM‐336372 (1‐5 μm) (Enzo Life Sciences, Farmingdale, NY, USA), or ERK inhibitor SCH772984 (1 μm) (AdooQ BioScience, Irvine, CA, USA) for 48 h. For DNA demethylation experiments, cells were treated with vehicle (0.1% ethanol) or 5 μm 5‐aza‐2′‐deoxycytidine (Sigma) for 48 h.
Hasegawa T., Adachi R., Iwakata H., Takeno T., Sato K, & Sakamaki T. (2017). ErbB2 signaling epigenetically suppresses microRNA‐205 transcription via Ras/Raf/MEK/ERK pathway in breast cancer. FEBS Open Bio, 7(8), 1154-1165.
+ Open protocol
+ Expand
5

Cellular Signaling Pathway Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
EGF was purchased from R&D Systems (Minneapolis, MN, USA). Phospho-ERK1/2, phospho-AKT (Thr308), phospho-AKT (Ser473), AKT, β-catenin, ERK1/2, and Histone3 antibodies were purchased from Cell Signaling Technology (Boston, MA, USA). MMP9 antibody was purchased from Bimake (Shanghai, China). GAPDH antibody and HRP-linked anti-rabbit secondary antibodies were purchased from Bioworld Technology (Louis Park, MN, USA). WNT7A antibody was purchased from Abcam (Cambridge, MA, USA). MEK inhibitor U0126 was purchased from Promega (Madison, WI, USA), and PI3K/AKT inhibitor LY294402 was purchased from Sigma (St. Louis, MO, USA).
Xie H., Ma Y., Li J., Chen H., Xie Y., Chen M., Zhao X., Tang S., Zhao S., Zhang Y., Du J., Zhang F, & Gu L. (2020). WNT7A Promotes EGF-Induced Migration of Oral Squamous Cell Carcinoma Cells by Activating β-Catenin/MMP9-Mediated Signaling. Frontiers in Pharmacology, 11, 98.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!