Chemicals, reagents & equipment Roche Diagnostics GmbH (Penzberg, Germany) produced all antibodies. They were aliquoted and stored at -80 • C. Murine monoclonal antibodies against human IgG and cynomolgus IgG were generated from mice immunized with purified human IgG or cynomolgus IgG. Antibodies from hybridoma supernatants were purified by protein A chromatography. Conjugates of cynomolgus monkey IgG with human IgG were generated in house [11] . Biotin (Bi)-and digoxigenin (Dig)-labeling was performed in house. Horse radish peroxidase (HRP)-labeled polyclonal anti-Dig antibody Fab fragments (pAb-Dig-S-Fab-HRP), phosphate-buffered saline (PBS), 2,2 -azinobis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS; ready to use solution) and Tween 20, were from Sigma Aldrich (MO, USA). Bovine plasma albumin and Roche Universal buffer were from Roche Diagnostics GmbH. Low Cross Buffer R was from Candor Bioscience GmbH, Wangen, Germany. Cynomolgus monkey serum (individual and pooled) was from Sera Laboratory International, Ltd (Haywords Heath, UK). Streptavidin (SA)-coated microtiter plates (SA-MTP) were from Microcoat Biotechnologie GmbH (Bernried, Germany). Antibodies and standards for the Ella cartridges were provided by Roche and labeled, immobilized and used by ProteinSimple, Biotechne, MN, USA, to produce cartridges for the two assays.
2 2 azino bis 3 ethylbenzthiazoline 6 sulfonic acid abts
Manufactured by Merck Group
Sourced in United States, Germany
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) is a water-soluble chromogenic compound. It is commonly used as a substrate in enzyme-linked immunosorbent assays (ELISA) for the detection and quantification of various analytes.
Automatically generated - may contain errors
Lab products found in correlation
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (5 mentions)
Ascorbic acid, by Merck Group (4 mentions)
Methanol, by Merck Group (4 mentions)
Acetonitrile, by Merck Group (3 mentions)
Tween 20, by Merck Group (2 mentions)
Folin ciocalteau reagent, by Merck Group (2 mentions)
Trolox, by Merck Group (2 mentions)
Sodium hydroxide (naoh), by Merck Group (2 mentions)
Ethanol, by Merck Group (2 mentions)
Acetic acid, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Bovine plasma albumin, by Roche (1 mentions)
Universal buffer, by Roche (1 mentions)
Silver nitrate, by Avantor (1 mentions)
Sh peg cooh, by JenKem Technology (1 mentions)
3 mercaptopropyl triethoxysilane mptes, by Merck Group (1 mentions)
Goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Sodium borohydride, by Merck Group (1 mentions)
Potassium phosphate monobasic, by Merck Group (1 mentions)
Trans rsv, by Merck Group (1 mentions)
17 protocols using 2 2 azino bis 3 ethylbenzthiazoline 6 sulfonic acid abts
1
Antibody Production and Characterization
2
Cardiac Troponin-I Biosensor Development
3
Niosomal Formulations for Antioxidant Delivery
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Tw80 was purchased to Riedel-de Haën (Munich, Germany) and Sp80 to Fluka (Buchs, Switzerland). Dod, hydrochloric acid, potassium chloride, sodium hydroxide, potassium phosphate monobasic and potassium persulfate were supplied by Merck (Darmstadt, Germany). 2,2’-Azinobis(3-ethylbenzthiazoline-6-sulfonic acid), (ABTS) ≥ 98%, trans-RSV ≥ 99%, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) ≥ 98% were provided by Sigma Aldrich (St. Louis, MO, USA). Ethanol absolute and methanol HPLC quality were obtained by Sintorgan (Buenos Aires, Argentina). MilliQ water was used in all experiments.
Niosomes formed by the mixture of Tw80 and Sp80 were named Tw80:Sp80 niosomes and that composed by the mixture of Tw80, Sp80 and Dod were called Tw80:Sp80:Dod niosomes.
Niosomes formed by the mixture of Tw80 and Sp80 were named Tw80:Sp80 niosomes and that composed by the mixture of Tw80, Sp80 and Dod were called Tw80:Sp80:Dod niosomes.
4
Comprehensive Phytochemical and Bioactive Analysis
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Chemicals and reagents: Folin-Ciocalteau reagent, sodium carbonate, aluminium trichlorue (AlCl3), gallic acid, ascorbic acid, quercetin, methanol, 2,2-diphenyl-picrylhydrazyl (DPPH), 2,2-azino-bis (3ethylbenzthiazoline-6-sulfonic acid) (ABTS), linoleic acid, sodium sulfate, sodium phosphate, soda and soybean lipoxygenase were purchased from SIGMA Aldrich (Spruce, St Louis, Germany); tween 80 from Sci-Tech Co. (Gangdong Guanghua, Shantou, China) ; Sabouroud chloramphenicol agar, Mueller Hinton broth, Mueller Hinton agar from OXOID (Basingstoke, HAMPSHIRE, ENGLAND). Phosphate Buffer Salin (PBS), Pancoll, Roswell Park Memorial Institute medium (RPMI 1640), Fetal Bovine Serum (FCS), Dimethylsulfoxide (DMSO) is from ThermoFisher (GIBCO, New York, USA).
5
Antioxidant Compounds Characterization
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Sodium dodecyl benzene sulfonate (SDBS, analytical purity) was acquired from Wuxi Ya Tai United Chemical Co., Ltd. (Jiangsu, PR China). 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) and 2,2′-azinobis-(3-ethyl benzthiazoline-6-sulfonic acid) (ABTS) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Ascorbic acid (Vc) and butylated hydroxy toluene (BHT) were obtained from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China).
6
Comprehensive Phenolic Standards Analysis
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In the LC–MS/MS analysis, the following compounds were used as standards: Alizarin (97%), acetohydroxamic acid (98%), butein (≥97.0%), caffeic acid (98%), catechin hydrate (99%), ellagic acid (≥96.0%), fumaric acid (99%), gallic acid (97.5%), quercetin (95%), oleuropein (98%), myricetin (96%), kaempferol (97%), luteolin (99%), naringenin (98%), phloridzin dehydrate (99%), rutin hydrate (94%), resveratrol (99%), salicylic acid (≥99.0%), 4-Hydroxybenzoic acid (99%), and vanillic acid (≥97.0%). These were purchased from Sigma Aldrich (Darmstadt, Germany). Stock solutions (1 mg/mL) were prepared from each phenolic standard in MeOH. Working solutions were prepared at 1 mg/L by taking appropriate portions from the stock solutions. The compounds used for the study (1,1-diphenyl-2-picryl-hydrazyl (DPPH), butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), α-tocopherol, ethanol, and HPLC-grade methanol) were obtained from Sigma (Sigma Aldrich, Darmstadt, Germany).
7
Phytochemical Analysis and Antioxidant Evaluation
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For the preparation of the extracts for HSCCC separation, the following analytical grade solvents were used: methanol, ethyl acetate, ethyl methyl ketone, MTBE (methyl tert-butyl ether), n-butanol, acetonitrile, and ethanol, which were acidified with either acetic acid, formic acid, or trifluoroacetic acid, all of which were purchased from Merck (Darmstadt, Germany).
For the HPLC–DAD–MS analyses, HPLC gradient grade methanol, acetic acid and water (Merck) were used. De-ionized water was purified by means of a Milli-Q system provided by Millipore (Bedford, MA, USA). The reference standards were purchased from Fluka (Buchs, Switzerland), Merck (Darmstadt, Germany), and Sigma (St. Louis, MO, USA).
To determine the extracts’ antioxidant activity, a saturated solution of Zn(CH3COO)2 (Panreac, Barcelona, Spain) and a solution of 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) (Sigma– Aldrich, Madrid, Spain) were mixed in phosphate buffer medium (pH 6). 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) supplied by Sigma–Aldrich was used for calibration.
For the HPLC–DAD–MS analyses, HPLC gradient grade methanol, acetic acid and water (Merck) were used. De-ionized water was purified by means of a Milli-Q system provided by Millipore (Bedford, MA, USA). The reference standards were purchased from Fluka (Buchs, Switzerland), Merck (Darmstadt, Germany), and Sigma (St. Louis, MO, USA).
To determine the extracts’ antioxidant activity, a saturated solution of Zn(CH3COO)2 (Panreac, Barcelona, Spain) and a solution of 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) (Sigma– Aldrich, Madrid, Spain) were mixed in phosphate buffer medium (pH 6). 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) supplied by Sigma–Aldrich was used for calibration.
8
Tyrosinase Inhibition Assay Protocol
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Arbutin, 2,2-diphenyl-1-picrylhydrazyl (DPPH), L-tyrosine, L-DOPA (L-3,4-dihydroxy phenylalanine), ascorbic acid, Tween-20, thiazolyl blue tetrazolium bromide (MTT), 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), O-tetradecanoyl phorbol-13-acetate (TPA), and mushroom tyrosinase were obtained from Sigma-Aldrich (St. Louis, MO). All other reagents and chemicals were of high-grade and acquired from commercial sources. The following antibodies were purchased from Bioworld Technology (St. Louis Park, MN, USA); anti-Tyr (Cat. No. BS6754), anti-TyrP-1 (Cat. No. sc-25543), anti-TyrP-2 (Cat. No. BS3320), anti-MITF (Cat. No. BS1550) for western blotting; or Cell Signaling Technology (Beverly, MA, USA); anti-phospho-JNK (Cat. No. 4668), anti-JNK (Cat. No. 9252), anti-phospho-p38 (Cat. No. 4511), anti-p38 (Cat. No. 9212), anti-p44/42MAPK (ERK1/2) (Cat. No. 9102), and anti-phospho-p44/42MAPK (ERK1/2) (Cat. No. 4376).
9
Bioactive Compound Extraction Protocol
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The materials used in this study were ethanol (HPLC grade), maltodextrin (Grain Processing Corporation, Muscatine, Schaumburg, IL USA), arabic gum (Frutarom, Schaumburg, IL, USA), orange essential oil (Sigma-Aldrich, St. Louis, MO, USA), soy lecithin (Sigma-Aldrich, USA), citric acid (Sigma-Aldrich, USA), calcium chloride (J.T. Baker, Phillipsburg, NJ, USA), commercial rennet (Christian Hansen, Mexico City, Mexico), tryptone–yeast extract agar (Sigma-Aldrich, USA), red bile violet agar (Sigma-Aldrich, USA), potato dextrose agar (Sigma-Aldrich, USA), Folin–Ciocalteau reagent (Sigma-Aldrich, USA), sodium carbonate (J.T. Baker, USA), sodium nitrite (J.T. Baker, USA), aluminum trichloride (Sigma-Aldrich, USA), sodium hydroxide (J.T. Baker, USA), 2,2-diphenyl-1-picrylhydrazyl (DPPH, Sigma-Aldrich, USA), methanol (analytical reagent grade), 2,2-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS, Sigma-Aldrich, USA), and potassium persulfate (analytical reagent grade).
10
Analytical Reagents for Scientific Research
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All chemicals were of analytical grade. Acetaminophen, sulfamethoxazole, and carbamazepine were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dextrose was purchased from Junsei (Tokyo, Japan). Potato Dextrose agar (PDA), malt extract, peptone, and yeast extract were purchased from BD Difco™ (Heidelgerg, Germany). Methanol and acetonitrile were purchased from J.T.Baker (Phillipsburg, NJ, USA), and 2, 2′-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and other chemicals were purchased from Sigma-Aldrich.
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