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5 protocols using 2 aminobenzaldehyde

1

Heterologous Expression of Human Organic Anion Transporter

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PLP, l-Orn, α-KG, 2-aminobenzaldehyde, dimethyl sulphoxide (DMSO), isopropyl-β-d-thiogalactoside (IPTG), phenylhydrazine hydrochloride and phenylmethylsulfonyl fluoride (PMSF) were purchased from Sigma. 1,8-Anilino-naphthalene sulfonic acid (ANS) was purchased from Molecular Probes. All other chemicals were of the highest purity available. The pYES2.1::hOAT expression plasmid coding for the complete cDNA of hOAT [22 (link)] was kindly provided by Prof. Leonardo Salviati of the University of Padova (Italy).
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2

Protein Labeling and Characterization

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L-Orn, α-KG, 2-aminobenzaldehyde, dimethyl sulphoxide, isopropyl-β-D-thiogalactoside, pyridoxine (PN), pyridoxamine (PM), pyridoxal (PL) were purchased from Sigma-Aldrich. 1,8-anilino-naphthalene sulfonic acid (ANS) was purchased from Molecular Probes. Growth media and additives were purchased from Gibco. PEGylated bis (sulfosuccinimidyl) suberate [BS (PEG) 5] was purchased from ThermoFisher. All other chemicals were of the highest purity available.
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3

Analytical Characterization of Organic Compounds

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Analytical grade chemicals, including 2-acetylpyrrole, 2-acetylpyridine, aniline, aniline hydrochloride, aniline sulfate, p-nitroaniline, p-nitroaniline hydrochloride, 2-aminobenzoic acid, 2-aminobenzonitrile, 2-aminobenzaldehyde, phenylhydrazine, phenylhydrazine hydrochloride, 2,4-dinitrophenylhydrazine, dicobalt octacarbonyl (Co2(CO)8), chromium hexacarbonyl (Cr(CO)6), 5% rhodium on an activated alumina (Rh/Al2O3) catalyst (99% purity) and internal standard 2,6-dimethylpyridine (2,6-DMP) were purchased from Sigma Aldrich, St. Louis, MO, USA. Silver nitrate (AgNO3) was purchased from POCH, Warsaw, Poland. Methanol (HPLC grade), dichloromethane (AR grade), hydrochloric acid (AR grade), toluene (AR grade) and sodium sulfate anhydrous (AR grade) were purchased from RCI Labscan, Bangkok, Thailand. Deionized water was obtained from a Nanopure Ultrapure water purification system (Thermo Fisher Scientific Inc., Waltham, MA, USA).
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4

Reagents for Dye Synthesis Protocols

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Reagents for dye synthesis were purchased from Alfa Aesar Co., Ward Hill, MA, USA (ninhydrin, 1,2-diaminobenzene) and Merck, Rahway, NJ, USA (ethyl bromoacetate, 2-indolinone, isatin, 1-methylisatin, 2-aminopyridine, 2-aminobenzaldehyde). Solvents used either for synthesis or spectroscopic measurements, i.e., acetic acid, isopropanol, ethanol, methanol, ethyl acetate, 1-methyl-2-pyrrolidinone (MP), chloroform, deuterated chloroform (CDCl3) and dimethyl sulfoxide (DMSO-d6) were provided by Merck. Monomer, trimethylolpropane triacrylate (TMPTA), commercial photoinitiator, camphorquinone (CQ) and electron donors: (phenylthio)acetic acid (PhTAA), phenylacetic acid (PhAA), 2-phenylpropionic acid (PhPA), N,N-dimethylaniline (DMA), 3-(phenylthio)acrylic acid (PhTACA) and hydrogen donor 2-mercaptobenzoxazole (MBX) were obtained from Sigma-Aldrich Co., St. Louis, MO, USA while N-phenyliminodiacetic acid (PhIAA) was from Lancaster, Kenilworth, NJ, USA. Thin layer chromatography plates (DC-Plastikfolien Silica gel 60 F254, 0.2 mm) from Merck Co. Rahway, NJ, USA.
The structures of the co-initiators used are shown in Figure 12. They are divided into two groups, i.e., electron donors and hydrogen atom donors.
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5

Enzymatic Assay Protocol

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L-Orn, α-KG, 2-aminobenzaldehyde, dimethyl sulphoxide, isopropyl-β-D-thiogalactoside, PLP, phenylmethylsulfonyl fluoride (PMSF) were purchased from Merck & Co (Rahway, NJ, USA), while 1,8-anilino-naphthalene sulfonic acid (ANS) was purchased from Molecular Probes (Eugene, OR, USA). All other chemicals were of the highest purity available.
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