Ag490

Manufactured by Bio-Techne

Sourced in United Kingdom

AG490 is a selective inhibitor of the Janus kinase 2 (JAK2) enzyme. It blocks the catalytic activity of JAK2, which is involved in the signal transduction pathways of various cytokines and growth factors.

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Lab products found in correlation

U0126, by Bio-Techne (3 mentions) Arecoline, by Merck Group (2 mentions) Pd153035, by Bio-Techne (2 mentions) Ly294002, by Bio-Techne (2 mentions) Aspirin, by Bio-Techne (1 mentions) Melatonin, by Merck Group (1 mentions) Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions) 5z 7 oxozeaenol, by Bio-Techne (1 mentions) P smad2 antibody, by Cell Signaling Technology (1 mentions) Sb431542, by Bio-Techne (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions) Antibiotic antimycotic solution, by Thermo Fisher Scientific (1 mentions) Y 27632, by Merck Group (1 mentions) Pkcζ pseudosubstrate, by Bio-Techne (1 mentions) Su6656, by Bio-Techne (1 mentions) G 6976, by Bio-Techne (1 mentions)

9 protocols using ag490

Alcoholic Hepatitis Murine Model

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At days 3, 6, and 11 of the modeling period, mice in the AH groups were gavaged a single dose of ethanol (31.5% ethanol(v/v), 400 μl/mouse), while mice in the control group were gavaged isocaloric dextrin maltose (45%(w/v), 400 μl/mouse). At day 10, MSCs with or without transfection were intraperitoneally (i.p.) administered to the AH+MSCs, AH+sc-MSCs, AH+siTSG-6-MSCs, and AH+MSCs+AG490 groups (5 × 10⁶ cells/mouse); following injection of MSCs, an additional dose of AG490 (Tocris, Bristol, UK) was given to the AH+MSCs+AG490 group (20 μg/mouse, i.p.); rmTSG-6 (R&D, Minneapolis, MN, USA) was injected to the AH+rmTSG-6 group (10 μg/mouse, i.p.); equal volumes of saline were administered to the control or AH group per protocol requirement. The gavage was always performed in the morning, and mice were then maintained on control or ethanol diet. After ethanol gavage, mice were lethargic and tachypneic, but they recovered within 4–6 h. The mice were always anesthetized 9 h post the last gavage with samples of blood, peritoneal fluid lavage, and liver tissues harvested for further analyses (Additional file 1).

Wan Y.M., Li Z.Q., Zhou Q., Liu C., Wang M.J., Wu H.X., Mu Y.Z., He Y.F., Zhang Y., Wu X.N., Li Y.H., Xu Z.Y., Wu H.M., Xu Y., Yang J.H, & Wang X.F. (2020). Mesenchymal stem cells alleviate liver injury induced by chronic-binge ethanol feeding in mice via release of TSG6 and suppression of STAT3 activation. Stem Cell Research & Therapy, 11, 24.

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Molecular Mechanisms Regulating Cell Signaling

Cited 2 times

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Dulbecco’s modified Eagles’ medium (DMEM), penicillin/streptomycin, trypsin/EDTA, fetal bovine serum (FBS), KGM-SFM medium, and phosphate-buffered saline (PBS) were from Gibco (Life Technologies, USA). Arecoline, melatonin and MTT were from Sigma (Sigma-Aldrich Chemical Company, St. Louis, MO, USA). ELISA kits for MMP-9 were from PeproTech (PeproTech, Inc., Rocky Hill, NJ, USA). Catalase, SB431542, 5Z-7-Oxozeaenol, PD153035, AG490, U0126, LY294002, and aspirin were purchased from Tocris. Phospho-TAK1 (p-TAK1) antibody was from Abcam (ab79583). Antibodies for TGF-β1, and GAPDH were from Santa Cruz. P-Smad2 antibody was from Cell Signaling Technology. PBL extract and ANE were prepared as before [8 (link), 12 (link), 13 (link), 21 (link)]. HC was synthesized as previously [22 (link)].

Chang M.C., Pan Y.H., Wu H.L., Lu Y.J., Liao W.C., Yeh C.Y., Lee J.J, & Jeng J.H. (2019). Stimulation of MMP-9 of oral epithelial cells by areca nut extract is related to TGF-β/Smad2-dependent and –independent pathways and prevented by betel leaf extract, hydroxychavicol and melatonin. Aging (Albany NY), 11(23), 11624-11639.

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Cell Culture and Signaling Inhibitors

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HUVECs were cultured in EBM2+ BulletKit (cells and media from Lonza, Rockville, MD). CHO-K1 cells were obtained from the American Type Culture Collection (Manassas, VA) and grown in high-glucose DMEM supplemented with 10% fetal bovine serum and antibiotic–antimycotic solution (all from Life Technologies/ThermoFisher Scientific, Grand Island, NY). Y-27632 (ROCK inhibitor) was purchased from Millipore (Billerica, MA). Cell-permeable C3 (RhoA inhibitor) transferase was purchased from Cytoskeleton (Denver, CO). U0126 (MEK inhibitor), LY294002 (PI3K inhibitor), Su6656 (Src family kinase inhibitor), AG 490 (JAK2 inhibitor), Gö6976 (PKCα inhibitor), FAK inhibitor 14, and PKCζ pseudosubstrate were from Tocris (Minneapolis, MN). All other reagents were from Sigma-Aldrich (St. Louis, MO) unless otherwise noted.

Scott D.W., Tolbert C.E, & Burridge K. (2016). Tension on JAM-A activates RhoA via GEF-H1 and p115 RhoGEF. Molecular Biology of the Cell, 27(9), 1420-1430.

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Immune Cell Profiling by Flow Cytometry

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For flow cytometry and sorting experiments, antibodies to CD4 (RM4-4), NK1.1 (PK136), CD49b (DX5), CD3 (17A2), Ly6G (1A8), B220 (RA3-6B2), CD11c (N418), CD45.2 (104), CD45.1 (A20), CX3CR1 (SA011F11), IA^b (AF6-120.1), CD11b (Mac1), CD90 (G7), lineage markers (17A2/RB6-8C5/RA3-6B2/Ter-119/M1/70), and CD127 (A7R34) were purchased from BioLegend. To stimulate neutrophils in vitro and for in vivo administration, we used either recombinant mouse IFN-λ2 (purchased from Peprotech) or recombinant mouse IFN-λ2 to which polyethylene glycol was attached (provided by Bristol-Myers Squibb). Recombinant mouse IFN-β was purchased from PBL interferonsource. Recombinant human IFN-λ2 and IFN-β were purchased from Peprotech. Puromycin was purchased from Sigma. Where indicated, specific chemical inhibitors were used. To inhibit protein synthesis, cycloheximide (Sigma) was used at a concentration of 10 μg/ml. To inhibit Jak signaling, pyridone 6 (BioVision) was used. To inhibit STAT-1 signaling, fludarabine (Tocris) was used. To inhibit Jak2 signaling, either AG490 (Tocris) or 1,2,3,4,5,6-hexabromocyclohexane (HBC, Tocris) was used. To stimulate neutrophils, either E. coli LPS (Serotype O55:B5 TLR grade, purchased from Enzo) or recombinant mouse TNF (Peprotech) was used.

Broggi A., Tan Y., Granucci F, & Zanoni I. (2017). IFN-λ suppresses intestinal inflammation by non-translational regulation of neutrophil function. Nature immunology, 18(10), 1084-1093.

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Leptin Signaling in Mouse HSCs

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The mouse HSC line (GRX) was maintained in DMEM, 5% FBS, and 1% penicillin/streptomycin as described [27 (link)]. Cells were treated with recombinant leptin (100 ng/ml; R&D systems, Minneapolis, MN) for 48 h. For neutralization of OPN, 40% confluent cells were treated with OPN-specific aptamers or biologically-inactive mutant sham aptamers (6.66 µg/ml). For PI3K inhibition, LY294002 (25 µM; Cell Signalling Technology, Danvers, MA) was applied to cells 30 min prior to treatment. AG490 (50 µM; Tocris Biosciences, Ellisville, MO) was used to inhibit JAK signaling [19 ].

Coombes J.D., Choi S.S., Swiderska-Syn M., Manka P.P., Reid D., Palma E., Briones-Orta M.A., Xie G., Younis R., Kitamura N., Peruta M.D., Bitencourt S., Dollé L., Oo Y.H., Mi Z., Kuo P.C., Williams R., Chokshi S., Canbay A., Claridge L.C., Eksteen B., Diehl A.M, & Syn W.K. (2015). Osteopontin is a proximal effector of leptin-mediated non-alcoholic steatohepatitis (NASH) fibrosis. Biochimica et biophysica acta, 1862(1), 135-144.

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Molecular Signaling in Alzheimer's Disease

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Anti-iNOS, anti-COX-2, anti-P-STAT3 (Tyr 705), anti-JAK2 and anti-P-JAK2 (Tyr1007/1008) antibodies were obtained from Cell Signaling (Danvers, MA). Anti-BACE1, anti-STAT3, anti-NF-κB p65, anti-IκBα, anti-SIRT1, and anti-histone H1 antibodies were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA). Anti-Aβ (6E10) anti-body was from Covance (Emeryville, CA). AG490 was from Tocris Bioscience (Bristol, UK). BAY11-7082, sirtinol, and quercetin were from Sigma-Aldrich (St. Louis, MO). Aβ_1–42 peptide was purchased from AnaSpec (Fremont, CA). Taxifolin and cilostazol were donated by Otsuka Pharmaceutical Co. Ltd. (Tokushima, Japan).

Park S.Y., Kim H.Y., Park H.J., Shin H.K., Hong K.W, & Kim C.D. (2016). Concurrent Treatment with Taxifolin and Cilostazol on the Lowering of β-Amyloid Accumulation and Neurotoxicity via the Suppression of P-JAK2/P-STAT3/NF-κB/BACE1 Signaling Pathways. PLoS ONE, 11(12), e0168286.

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Keratinocyte Growth and Inflammation Modulation

Cited 1 time

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Keratinocyte growth medium (KGM-SFM), pituitary gland extract and EGF were purchased from Gibco (Life Technologies, BRL, Grand Island, NY, USA). Arecoline, catalase, aspirin and 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) were obtained from Sigma (Sigma Chemical Company, St. Louis, MO, USA). ANE was prepared and weighed as previously described [25 (link), 32 (link)]. PGE₂ and 8-isoprostane (8-iso-PGF_2α) Enzyme-linked immunosorbant assay (ELISA) kits were obtained from Cayman Chemical Company (Ann Arbor, MI, USA). Human IL-1α ELISA kits were from PeproTech (Rocky Hill, NJ, USA), whereas ADAM17 and EGF ELISA kits were from R&D Systems (Minneapolis, MN, USA). GM6001, PD153035, AG490, pp2, U0126, α-naphthoflavone and ZnPP were obtained from Tocris or Cayman. Anti-EGF, anti-IL-1α and anti-TNFα neutralizing aby were obtained from PeproTech. Antibodies against COX-2, cdc2, ADAM9 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were from Santa Cruz, whereas antibodies for cytokeratin 5 and cytokeratin 14 were from GeneTex (GeneTex International Corporation, Global, Hsin-Chu, Taiwan). IRAK-1/4 inhibitor was from APExBIO (Houston, TX, USA).

Chang M.C., Chan C.P., Chen Y.J., Hsien H.C., Chang Y.C., Yeung S.Y., Jeng P.Y., Cheng R.H., Hahn L.J, & Jeng J.H. (2016). Areca nut components stimulate ADAM17, IL-1α, PGE2 and 8-isoprostane production in oral keratinocyte: role of reactive oxygen species, EGF and JAK signaling. Oncotarget, 7(13), 16879-16894.

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Sheep Prolactin Electrophysiology Protocol

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Sheep PRL and picrotoxin (PTX) were purchased from Sigma (St Louis, MO) and AG490 from Tocris Cookson (Bristol, UK). Drugs were prepared as stock solutions, stored frozen in the dark, and diluted to final concentration immediately before use. Sheep PRL (44.5 μM) was prepared in milli‐Q water while AG490 (100 mM) and 17β‐estradiol (50 mg/ml) were prepared in dimethyl sulfoxide and dissolved in sesame oil for s.c. injection. Drugs were dissolved in oxygenated aCSF at the desired concentration immediately before use. Slices were incubated (20 min) in sheep PRL before being transferred to the immersion chamber, in which PRL was present for the entire recording period at the same concentration.

Zamora‐Moratalla A, & Martín E.D. (2020). Prolactin enhances hippocampal synaptic plasticity in female mice of reproductive age. Hippocampus, 31(3), 281-293.

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JAK2 Inhibition Regulates IL-6-Induced Feeding

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ICV injection of AG490 (50 mM) (cat #0414, Tocris Bioscience, Bristol, United Kingdom) was used to inhibit JAK2 as previously described (Damm et al., 2013 (link); Quaresma et al., 2017 (link)). ICV injection of AG490 or its vehicle was performed twice daily in the afternoon; 30 min later, we injected IL-6 ICV before the lights were turned off. Food intake measurements or euthanasia started after 06:30 p.m. (dark cycle). For this experiment, we used wt/wt mice (6–8 weeks of age) obtained from the B6-tub/tub mouse colony.

de Oliveira Micheletti T., Cassia dos Santos A., Rocha G.Z., Silva V.R., Quaresma P.G., Assalin H.B., Junqueira F.S., Ropelle E.R., Oliveira A.G., Saad M.J, & Prada P.D. (2022). Acute exercise reduces feeding by activating IL-6/Tubby axis in the mouse hypothalamus. Frontiers in Physiology, 13, 956116.

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