D-glucose, acetonitrile (Sigma, USA), NaCl (Vekton, Russia), fluorescent dyes BADAN and quinine sulfate (AnaSpec, USA), guanidine hydrochloride (Nacalai Tesque, Japan), and glycerol (Merck, Germany) were used without further purification. The mutant form of the D-glucose/D-galactose–binding protein GGBP/H152C was obtained, separated and purified as described previously (Stepanenko et al., 2011a (link)). The labeling of GGBP/H152C with the fluorescent dye BADAN was performed as described by Khan, Saxl & Pickup (2010) (link). The experiments were performed in solutions with protein concentrations ranging from 0.2 to 0.8 mg/ml. For the formation of the protein-ligand complex, 5 µM–20 mM of D-glucose was added to the protein solution. The measurements were made in buffer solutions containing citric acid and Na2HPO4 (pH 2.8, 4.2, 6.0, 7.1), PBS (pH 7.4) and TrisHCl (pH 7.2, 9.6).
Guanidine hydrochloride
Manufactured by Nacalai Tesque
Sourced in United States, Japan
Guanidine hydrochloride is a chemical compound commonly used in laboratory settings. It is a crystalline solid that is soluble in water and functions as a chaotropic agent, capable of disrupting the structure of proteins and nucleic acids.
Automatically generated - may contain errors
Lab products found in correlation
Acetonitrile, by Merck Group (1 mentions)
Glycerol, by Merck Group (1 mentions)
Monoiodoacetate, by Merck Group (1 mentions)
N glycosidase f, by Roche (1 mentions)
Dithiothreitol (dtt), by Merck Group (1 mentions)
Ultrapure tris, by Thermo Fisher Scientific (1 mentions)
Sodium acrylate, by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Nacalai Tesque (1 mentions)
Triton x 100, by Nacalai Tesque (1 mentions)
N n methylene bisacrylamide, by Nacalai Tesque (1 mentions)
Acrylamide aa, by Merck Group (1 mentions)
Proteinase k, by Nacalai Tesque (1 mentions)
Seaplaque agarose, by Lonza (1 mentions)
Glycerol, by Fujifilm (1 mentions)
Sucrose, by Fujifilm (1 mentions)
Glucose, by Nacalai Tesque (1 mentions)
Human hemoglobin, by Merck Group (1 mentions)
Glycine, by Nacalai Tesque (1 mentions)
Betaine, by Fujifilm (1 mentions)
Maltose, by Nacalai Tesque (1 mentions)
4 protocols using guanidine hydrochloride
1
Fluorescent Labeling of Glucose-Binding Protein
2
N-Glycosidase F Purification Protocol
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N-Glycosidase F was purchased from Roche Diagnostics (Mannheim, Germany). Guanidine hydrochloride was purchased from Nacalai Tesque (Kyoto, Japan). Dithiothreitol and monoiodoacetate were purchased from Sigma (St. Louis, MO, USA). All other reagents were of the highest quality available.
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N-Glycosidase F was purchased from Roche Diagnostics (Mannheim, Germany). Guanidine hydrochloride was purchased from Nacalai Tesque (Kyoto, Japan). Dithiothreitol and monoiodoacetate were purchased from Sigma (St. Louis, MO, USA). All other reagents were of the highest quality available.
3
Hydrogel-Based Cell Encapsulation Protocol
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Antibodies used in this study are listed in Supplemental Table S1. The following reagents were purchased from the manufacturers as noted: sodium acrylate (SA) (#408220) from Sigma-Aldrich; acrylamide (AA) (#00809-85), N,N′-methylenebisacrylamide (#22402-02), ammonium persulfate (APS) (#02627-34), N,N,N′,N′-tetramethylethylenediamine (TEMED) (#33401-72), guanidine hydrochloride (#17318-95), EDTA (#15105-35), Triton X-100 (#12967-45), and proteinase K (#29442-14) from Nacalai Tesque; 6-((acryloyl)amino)hexanoic acid, succinimidyl ester (Acryloyl-X, SE) (#A20770), and UltraPure Tris (#15504-020) from Invitrogen; and SeaPlaque Agarose (#50100) from Lonza.
The monomer solution containing 8.6 g/100 ml of SA, 2.5 g/100 ml of AA, 0.15 g/100 ml of N,N′-methylenebisacrylamide, 11.7 g/100 ml of sodium chloride, and 1× phosphate-buffered saline (PBS) was prepared and stored at 4°C until use. Digestion buffer containing 50 mM Tris-HCl (pH 8.0), 1 mM EDTA, 0.5% Triton X-100, and 0.8 M guanidine hydrochloride was prepared and stored at 4°C until use.
The following materials were purchased from the manufacturers: Grace Bio-Labs CultureWell removable chambered cover glass (Sigma-Aldrich; #GBL112358), six-well glass-bottom plates with high-performance #1.5 cover glass (Cellvis; P06-1.5H-N), and coverslips (9-mm diameter) (Matsunami Glass; #CS01029).
The monomer solution containing 8.6 g/100 ml of SA, 2.5 g/100 ml of AA, 0.15 g/100 ml of N,N′-methylenebisacrylamide, 11.7 g/100 ml of sodium chloride, and 1× phosphate-buffered saline (PBS) was prepared and stored at 4°C until use. Digestion buffer containing 50 mM Tris-HCl (pH 8.0), 1 mM EDTA, 0.5% Triton X-100, and 0.8 M guanidine hydrochloride was prepared and stored at 4°C until use.
The following materials were purchased from the manufacturers: Grace Bio-Labs CultureWell removable chambered cover glass (Sigma-Aldrich; #GBL112358), six-well glass-bottom plates with high-performance #1.5 cover glass (Cellvis; P06-1.5H-N), and coverslips (9-mm diameter) (Matsunami Glass; #CS01029).
4
Characterization of Biomolecular Interactions
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NaH 2 PO 4 , Na 2 HPO 4 , levulose, xylose, sucrose, glycerol, betaine, D-sorbitol, and lactobionic acid were purchased from Wako Chemical (Japan). Human hemoglobin was purchased from Sigma-Aldrich. Guanidine hydrochloride, glucose, mannose, galactose, fucose, maltose, and glycine were purchased from Nacalai Tesque. Trehalose and glucosamine hydrochloride were purchased from Tokyo Chemical Industry. L-proline was purchased from Peptide Institute, Inc. (Japan). L-arginine hydrochloride was purchased from Kishida Chemical. UV-Vis spectra were recorded using a JASCO V-530 spectrometer with a bandwidth of 2.0 nm, data acquisition interval of 1 nm, and a range of 300-500 nm.
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