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2 protocols using 4 benzoylbenzoic acid

1

Synthesis of Norbornene-Functionalized PDMS Polymers

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N-(hexanoic acid)-cis-5-norbornene-exo-dicarboximide was prepared according to literature (59 ). Grubbs’ second-generation metathesis catalyst [(H2IMes)(PCy3)(Cl)2Ru = CHPh] was generously provided by Materia. Grubbs’ third-generation metathesis catalyst [(H2IMes)(pyr)2(Cl)2Ru = CHPh] (G3) was prepared according to literature (60 ). Methanol (purity of >99.8%; Fisher Scientific, A412), dichloromethane (purity of >99.5%; Fisher Scientific, D37), toluene (purity of >99.5%; Fisher Scientific, T324-500), triethylamine (purity of >99%; Fisher Scientific, ACROS Organics, AC15791), and ethyl vinyl ether (purity of >99%; Fisher Scientific, ACROS Organics, AC119082500) were used as received. CDCl3 (99.8%) was purchased from Cambridge Isotope Laboratories (DLM7) and used as received. Bis(hydroxyalkyl)-terminated PDMS (Sigma-Aldrich, 481246), hydroxyalkyl-terminated PDMS (Gelest, MCR C18 and MCR C22), polyethylene glycol monomethyl ether 550 (TCI, P2184), succinic anhydride (purity of >99%; Sigma-Aldrich, 239690), 4-benzoylbenzoic acid (purity of 99%; Sigma-Aldrich, B12407), N,N-dimethylamino pyridine (purity of 99%; Alfa Aesar, H51715), and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (purity of 99%; Oakwood Chemical, 024810) were used as received.
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2

In Vitro CK2 Kinase Assay

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ATP was obtained from Fisher. CK2 was purchased from New England Biolabs. CK2 peptide substrate (RRREEETEEE) was purchased from Promega. The 2,2′-ethylenedioxy bis-(ethylamine), alpha-cyano-4-hydroxy cinnamic acid, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) were purchased from Acros. The 4-benzoyl benzoic acid, N-methyl morpholine (NMM), and 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES) were obtained from Sigma–Aldrich. Tris-base and sodium chloride were obtained from JT Baker. D4-MeOH, D2O, and CDCl3 were purchased from Cambridge Isotope Lab. Inc. Flash chromatography was performed on silica gel, 200–400 mesh (Merck). A-25 sephadex was purchased from Aldrich. The anti-CK2 antibody was obtained from Millipore and Sigma–Aldrich. Goat anti-Mouse IgG H&L (HRP) secondary antibody was purchased from Abcam. SuperSignal West Dura Chemiluminescent Substrate was purchased from Thermo Scientific. Coomassie blue stain was purchased from NuSep.
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