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4 protocols using l glycine

1

Taste Receptor Activation by Amino Acids

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Agar (purified powder) was obtained from Sigma-Aldrich (St Louis, MO, USA). As tastants, D-fructose (CAS: 57-48-7, Wako Pure Chemical Industries, Ltd., Osaka, Japan), as well as L-alanine (ALA; CAS: 56-41-7), L-arginine monohydrochloride (ARG; CAS: 1119-34-2), L-asparagine monohydrate (ASN; CAS: 5794-13-8), L-aspartic acid (ASP; CAS: 56-84-8), L-cysteine monohydrochloride hydrate (CYS; CAS: 7048-04-6), L-histidine monohydrochloride (HIS; CAS: 645-35-2), L-isoleucine (ILE; CAS: 73-32-5), L-leucine (LEU; CAS: 61-90-5), L-lysine monohydrochloride (LYS; CAS: 657-27-2), L-glutamine (GLN; CAS: 56-85-9), L-glutamic acid (GLU; CAS: 56-86-0), L-glycine (GLY; CAS: 56-40-6), L-methionine (MET; CAS: 63-68-3), L-phenylalanine (PHE; CAS: 63-91-2), L-proline (PRO; CAS: 147-85-3), L-serine (SER; CAS: 56-45-1), L-threonine (THR; CAS: 72-19-5), L-tryptophan (TRP; CAS: 73-22-3), L-tyrosine (TYR; CAS: 60-18-4) and L-valine (VAL; CAS: 72-18-4) were used at concentrations of 10 mmol l−1 (TYR was used in 0.5 mmol l−1 due to low solubility). Amino acids were obtained from Wako Pure Chemical Industries, Ltd., Osaka, Japan (TYR, SER, CYS, VAL), Sigma-Aldrich, St Louis, MO, USA (ILE, PRO) or Nacalai Tesque, Kyoto, Japan (ALA, ARG, ASN, ASP, LEU, LYS, GLN, GLY, MET, PHE, THR, TRP, TYR).
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2

Synthesis and Analysis of Amino Acids

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L-Kynurenine sulfate salt, KYNA, and L-tyrosine disodium salt hydrate were purchased from Sigma Chemical Co. (St. Louis, MO, USA). L-Alanine, L-arginine, L-asparagine, L-aspartate, L-cysteine hydrochloride monohydrate, L-glutamine, L-glutamate, L-glycine, L-histidine, L-isoleucine, L-leucine hydrochloride, L-lysine, L-methionine, L-phenylalanine, L-proline, L-serine, L-threonine, and L-valine were purchased from Wako Pure Chemical Industries (Osaka, Japan). Rodent diet (MF) was obtained from Oriental Yeast Co., Ltd (Tokyo, Japan). All other chemicals were of the highest commercially available purity.
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3

Analytical-Grade Chemical Compounds

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Propionic acid (PA), butyric acid (BA), L-glycine (Gly), L-alanine (Ala), L-serine (Ser), L-threonine (Thr), L-cysteine (Cys), L-methionine (Met), L-valine (Val), L-leucine (Leu), L-isoleucine (Ile), L-phenylalanine (Phe), L-tryptophan (Trp), L-aspartic acid (Asp), L-glutamic acid (Glu), L-asparagine (Asn), L-glutamine (Glu), L-histidine (His), L-asparagine monohydrate (Arg), L-tyrosine (Tyr), and L-ascorbic acid were purchased from FUJIFILM Wako (Osaka, Japan). Acetic acid (AA), L-lysine (Lys), and sodium butyrate were purchased from Nacalai Tesque (Kyoto, Japan), and proline (Pro) was purchased from Sigma-Aldrich (St. Louis, MO, USA). All chemicals were of analytical grade.
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4

Characterization of Modified Human Serum Albumin

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Succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC) was purchased from Tokyo Chemical Industry Co. Ltd. Human serum albumin (HSA) was purchased from the Japan Blood Products Organization. Bis(3,5-dibromosalicyl)fumarate (DBBF) was purchased from LKT Laboratories Inc. L-Glycine was purchased from Wako Pure Chemical Industries Ltd. Inositol hexaphosphate (IHP) was purchased from Sigma-Aldrich Corp. Human adult Hb (HbA) was purified from red blood cell concentrates received from the Japanese Red Cross Society (see ESI †). Other chemicals of special grade were used without further purification unless otherwise noted. Water was deionized (18.2 MO cm) using water purification systems (Elix UV and Milli Q Reference; Millipore Corp.). Native-PAGE was performed using an electrophoresis power supply (EPS 301; GE Healthcare) using 5-12% polyacrylamide precast gradient gel (SuperSep Ace 5-12%; Wako Pure Chemical Industries Ltd). Isoelectric focusing (IEF) was done using an electrophoresis power supply (EPS 601; GE Healthcare) using a pH 3-10 IEF gel (Novex; Invitrogen Corp.). A GE Healthcare IEF calibration kit Broad pI (pH 3-10) was used for the protein marker. The voltage was raised gradually to 500 V for 2 h.
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