Lutein (purity > 90%), alcalase (enzyme activity > 200 U/mg), bile salts, and molecular weight standards including bacitracin, aprotinin, myohemoglobin, bovine serum albumin, and thyroglobulin were purchased from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China). Soybeans were bought from Fengyuan Zhongye Co., Ltd. (Lianyungang, China). Pepsin (enzyme activity > 2500 U/mg), trypsin (enzyme activity > 2500 U/mg), 8-aniline-1-naphthalensulfonic acid (ANS), and trinitrobenzenesulfonic acid (TNBS) were purchased from Sigma-Aldrich Co., (St. Louis, MO, USA). All other reagents and solvents were of at least analytical grade.
Trinitrobenzene sulfonic acid tnbs
Manufactured by Merck Group
Sourced in United States
Trinitrobenzene sulfonic acid (TNBS) is a chemical compound used in various laboratory applications. It is a crystalline solid with a yellowish-brown appearance. TNBS is commonly employed as a reagent for the detection and quantification of primary amines in biochemical and analytical procedures.
Automatically generated - may contain errors
Lab products found in correlation
Trypsin, by Merck Group (1 mentions)
Pepsin, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Yuanye Bio-Technology (1 mentions)
Lutein, by Yuanye Bio-Technology (1 mentions)
Bacitracin, by Yuanye Bio-Technology (1 mentions)
Aprotinin, by Yuanye Bio-Technology (1 mentions)
Alcalase, by Yuanye Bio-Technology (1 mentions)
Am630, by Bio-Techne (1 mentions)
O 1918, by Bio-Techne (1 mentions)
Am251, by Bio-Techne (1 mentions)
Abn cbd, by Bio-Techne (1 mentions)
1 ethyl 3 3 dimethylaminopropyl carbodiimide edc, by Merck Group (1 mentions)
2 n morpholino ethanesulfonic acid mes, by Merck Group (1 mentions)
Nitric acid, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Ethanol, by Merck Group (1 mentions)
Glutaraldehyde, by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Sodium sulfate na2so4, by Merck Group (1 mentions)
17 protocols using trinitrobenzene sulfonic acid tnbs
1
Soy Protein Extraction and Characterization
2
Pharmacological Modulation of TNBS Colitis
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Trinitrobenzene sulfonic acid (TNBS) was purchased from Sigma-Aldrich (Oakville, Ontario, Canada). Abn-CBD, dissolved in methyl acetate, O-1918 (1,3-Dimethoxy-5-methyl-2-[(1R,6R)-3-methyl-6-(1-methylethenyl)-2-cyclohexen-1-yl]benz-ene), AM630 (6-iodo-2-methyl-1-[2-(4-morpholinyl)ethyl-1H-indol-3-yl](4-methoxyphenyl)methanone) and AM251 (N-(Piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide) were obtained from Tocris Bioscience (Bristol, UK). Because of its toxicity, methyl acetate was evaporated prior to the in vivo experiments and ethanol was used instead as a solvent. Abn-CBD was then further diluted in Tween 80 (10 %) and sterile saline. Vehicle consisted of ethanol, Tween 80 and sterile saline (1:1:8). AM630 and AM251 were dissolved in dimethyl sulfoxide (DMSO, 99.7 %) and further diluted with vehicle. 45 min prior to the induction of TNBS colitis, mice were injected intraperitoneally (i.p.) with 5 mg/kg AM630, AM251, O-1918 or vehicle, followed by 5 mg/kg Abn-CBD, 15 min later. As a single dose of Abn-CBD was ineffective (preliminary data not shown), mice were injected twice daily for 3 days. For in vitro assays, 10 mM stock solutions (in DMSO) of Abn-CBD and the CB receptor antagonists were prepared.
3
Functionalization of Biomolecules with NHS
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N-Hydroxysuccinimide (NHS) was obtained from Fluka Chemie AG, Buchs, Switzerland. Acetic acid, ammonium chloride (NH4Cl), ammonium sulfate ((NH4)2SO4), bovine serum albumin, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), ethanol, glutaraldehyde, hydrogen chloride, nitric acid, paraformaldehyde, sodium chloride (NaCl), sodium hydrogen phosphate (Na2HPO4) and sodium sulfate (Na2SO4) were from Merck, Darmstadt, Germany. Glycine was from Scharlau Chemicals, Barcelona, Spain. Barium perchlorate (Ba(ClO4)2), barium sulfate (BaSO4), calcium chloride (CaCl2), calcium sulfate (CaSO4), magnesium chloride (MgCl2), magnesium perchlorate (Mg(ClO4)2), 2-(N-morpholino)ethane sulfonic acid (MES) and trinitrobenzene sulfonic acid (TNBS) were from Sigma Aldrich, St. Louis, MO, USA. Barium chloride (BaCl2) and magnesium sulfate (MgSO4) were from VWR International, Radnor, PA, USA.
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N-Hydroxysuccinimide (NHS) was obtained from Fluka Chemie AG, Buchs, Switzerland. Acetic acid, ammonium chloride (NH4Cl), ammonium sulfate ((NH4)2SO4), bovine serum albumin, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), ethanol, glutaraldehyde, hydrogen chloride, nitric acid, paraformaldehyde, sodium chloride (NaCl), sodium hydrogen phosphate (Na2HPO4) and sodium sulfate (Na2SO4) were from Merck, Darmstadt, Germany. Glycine was from Scharlau Chemicals, Barcelona, Spain. Barium perchlorate (Ba(ClO4)2), barium sulfate (BaSO4), calcium chloride (CaCl2), calcium sulfate (CaSO4), magnesium chloride (MgCl2), magnesium perchlorate (Mg(ClO4)2), 2-(N-morpholino)ethane sulfonic acid (MES) and trinitrobenzene sulfonic acid (TNBS) were from Sigma Aldrich, St. Louis, MO, USA. Barium chloride (BaCl2) and magnesium sulfate (MgSO4) were from VWR International, Radnor, PA, USA.
4
Lupine Protein Isolate Production
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Lupine protein isolate (LPI) with protein content of 85% was produced according to the alkalization and precipitation method (Rezvankhah et al., 2021a (link)). Alcalase as an endopeptidase (the commercially called Alcalase 2.4 L), company reported activity of 2.4 anson units per gram (AU/g) with a density of 1.18 g/mL that has been originated from Bacillus licheniformis, and Flavourzyme as an exopeptidase (the commercially called Flavourzyme 1000 L), company reported activity of 1000 leucine amino peptidase units per gram (LAPU/g) with a density of 1.28 g/mL that has been originated from Aspergillus oryzae were provided from Novozymes Co. (Bagsværd outside of Copenhagen). Alcalase and Flavourzyme were used to produce peptides or single amino acids. Microbial transglutaminase (MTGase) with the activity of 100 U/g was prepared from Ajinomoto Co. Gum Arabic (GA) was provided from Ingredion Co. Trinitrobenzenesulfonic acid (TNBS) was purchased from Sigma Company. All other chemicals used were of analytical and HPLC (high performance liquid chromatography) grade.
5
Silk Fibers Functionalization and Evaluation
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A. pernyi silk fibers were purchased from Liaoning Province (People’s Republic of China). PEI (branched; molecular weight: 25 kDa), EDC, 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT), ethylene-diamine, and trinitrobenzene sulfonic acid (TNBS) were purchased from Sigma-Aldrich (St Louis, MO, USA). Plasmid DNA encoding enhanced green fluorescent protein (EGFP; 4,733 bp; Genbank accession number: U55762) was purchased from Clontech (Mountain View, CA, USA), amplified in the DH5α Escherichia coli strain, and purified using a Qiagen Plasmid Mega Kit (Qiagen, Chatsworth, CA, USA). The concentration of plasmid DNA was determined by measuring the ultraviolet (UV) absorbance at 260 nm. The human endothelial cell line EA.hy926 (CRL-2922™) was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The experiments were formally reviewed and approved by the ethics committee of Soochow University. Reviewed by the ethics committee, the experimental design and implementation have fully considered the principle of security; the experimental content did not exist potential damage and risk, and followed the principles outlined in the Declaration of Helsinki.
6
Yak Bone Peptides Extraction Optimization
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Yak bones (leg bone) were obtained from the Tibet Academy of Agricultural and Animal Husbandry Sciences (Tibet, China). The bones were mechanically ground into granules with a diameter ranging from 3–5 mm and stored at −80 °C until analyzed. Pepsin (EC number 3.4.23.1, 1200 U mg−1), trypsin (EC number 3.4.21.4, 92 383 U mg−1), Alcalase (EC number 3.4.21.63, 221 756 U mg−1), Flavourzyme (EC number 3.4.11.1, 15 311 U mg−1), Protamex (56 084 U mg−1), Neutrase (EC number 3.4.24.28, 19 267 U mg−1) and papain (EC number 3.4.22.2, 37 018 U mg−1) were purchased from Solarbio (Beijing, China). Soluble peptides content was measured using a bicinchoninic acid (BCA) Protein Assay Kit from Beyotime (Shanghai, China). Trifluoroacetic acid (TFA), sodium dodecyl sulfate (SDS), and trinitrobenzene sulfonic acid (TNBS) were purchased from Sigma-Aldrich (Shanghai, China). HPLC grade acetonitrile (ACN) was purchased from Thermo Fisher Scientific (Waltham, USA). GP-16 and GD-18 (purity > 98%) was chemical synthesized and purified using high performance liquid chromatography (HPLC) by Beijing Protein Innovation (Beijing, China). All other chemicals and reagents were of analytical grade, unless otherwise stated.
7
Quantifying Cross-Linking Degree in Fish Skin Scaffolds
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The cross‐linking degree in the fish skin scaffolds has been achieved by assessing the free amine content following the method conducted by Ma and colleagues.27 The samples (3–5 mg) were socked in a solution containing 1 mL of 4% (w/v) NaHCO3 (Merck‐Millipore, USA) and 1 mL of 0.5% (v/v) tri‐nitrobenzene sulfonic acid (TNBS) (Sigma‐Aldrich, USA) for 2 hours at 40°C. Then, 3 mL of 6 moL/L HCl (Merck‐ Millipore, USA) was added to the mixture and heated for 90 min at 60°C. The solution was diluted with distilled water, and then the absorbance was measured at 345 nm with a UV spectrophotometer (Shimadzu 160‐UV, Tokyo; Japan). The content of the free amine group was calculated using Equation (1 ) as follows.33 , 35 where “N” is the free amine group content (mol·g−1), “A” is the absorbance, “V” is the volume of solution (mL), “ε” = 14.600 (mL·mmol−1·cm−1), “l” is path length (cm), and “m” is the weight of the sample (mg). Then, the free amine group content (N) was used to determine the cross‐linking degree (D). The calculation was done using the following equation (Equation 2 ):
8
TRPC4/5 Antagonist HC-070 in Rat Pain Models
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HC-070 was synthesized by Orion Pharma (Espoo, Finland) and used as a TRPC4/5 channel antagonist at an oral dose range of 3–10 mg/kg in rat neuropathic and visceral pain models. HC-070 was dissolved in a suspension of 0.5% (w/v) methyl cellulose (MC) in purified water. Morphine was purchased from Francopia (Catalog reference 3695, Batch No. KR00017 or JR00002, Paris, France), dissolved in 0.9% NaCl solution, and used at a subcutaneous (s.c.) dose of 3 mg/kg in the rat neuropathic pain model or a dose of 1 mg/kg in the model of partial restraint stress in female rats. Trinitrobenzene sulfonic acid (TNBS), purchased from Sigma-Aldrich (Catalog reference 92822, Batch No. BCBT6864, Chesnes, France), solution (1 mL/kg) was prepared in 25% Ethanol Absolut. The κ-opioid agonist (−)U-50,488H, as the positive reference, was purchased from Sigma-Aldrich (Catalog reference U111, Batch No. 070M4626V), dissolved in 0.9% NaCl solution, and used at a subcutaneous (s.c.) dose of 3 mg/kg in the rat TNBS model.
9
Rat Model of Colitis with TNBS
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Sixty Sprague–Dawley (SD) rats (equal ratio of male and female), weighing 250 ± 20 g, were obtained from Experimental Animal Center of Nanjing University (Nanjing, China). Sulphasalazine (SASP) (250 mg) was purchased from the National Institutes for Food and Drug Control (Beijing, China). Trinitro-benzene-sulfonic acid (TNBS) was purchased from Sigma Chemical (St. Louis, MO, USA).
10
Rat Model of Inflammatory Bowel Disease
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PCT was obtained from Tokyo Chemical Industry (Tokyo, Japan). Trinitrobenzene sulfonic acid (TNBS) was purchased from Sigma Chemical Co. (Perth, Australia). All other chemicals were reagent-grade, commercially available products. Ten-week-old male male Sprague Dawley rats (Samtako Bio Korea, Osansi, Republic of Korea) were housed in the university animal facility with controlled temperature, humidity, and dark/light cycle. The animal protocol used in this study has been reviewed and approved by the Pusan National University-Institutional Animal Care and Use Committee based on their ethical procedures and scientific care.
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