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2 protocols using 2 2 amino 3 methoxyphenyl 4h 1 benzopyran 4 one pd98059

1

Fura-2-AM and DiSC3(5) Calcium Signaling

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Fura-2-acetoxymethyl ester (Fura-2-AM) and 3’,3’,3’-dipropylthiadicarbocyanine iodide [DiSC3(5)] were purchased from Invitrogen (Carlsbad, CA, USA). ω-conotoxin MVIIC, rottlerin, 2-[1-(3-dimethylaminopropyl)indol-3-yl]-3-(indol-3-yl) maleimide (GF109203X), 5,6,7,13-tetrahydro-13-methyl-5-oxo-12H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-12-propanenitrile (Go6976) and N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide (H89) were purchased from Tocris Bioscience (Bristol, UK). Echinacoside, dantrolene, dl-threo-beta-benzyl-oxyaspartate (dl-TBOA), 7-chloro-5-(2-chloropheny)-1,5-dihydro-4,1-benzothiazepin-2(3H)-one (CGP37157), 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one) (PD98059), ethylene glycol bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid (EGTA) and all other reagents were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA).
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2

Antibody Immunoblot Reagents Protocol

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Monoclonal antibodies against β-tubulin, Nur77 and FLAG M2 were purchased from Millipore Corporation (Billerica, MA, USA), Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA) and Sigma (St. Louis, MO, USA), respectively; polyclonal antibodies against phospho-ERK1/2 (pERK1/2) and total ERK1/2 were purchased from New England Biolabs, Inc. (Beverly, MA, USA); horseradish peroxidaseconjugated goat-anti-rabbit, goat-anti-mouse secondary antibodies, as well as immuno-blot polyvinylidene fluoride membrane were purchased from Bio-Rad Laboratories (Hercules, CA, USA); 2-(2amino-3-methoxyphenyl) 4H-1-benzopyran-4-one (PD98059) and 8-bromo-cAMP (8Br-cAMP) were purchased from Sigma (St. Louis, MO, USA).
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