Gallic acid and folin ciocalteu as well as chemicals for ORAC assay such as sodium fluorescein, 2,2′-azobis(2-amidinopropane)dihydrochloride (AAPH), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox®), Nunc Micro-well™ plates, phosphate buffer (pH 7.4) were purchased from Sigma Aldrich. ORAC experiment was performed on fluorometer, FLUOstar OPTIMA, Franka Ganske, BMG LABTECH, Offenburg, Germany. DPPH free radical solution, vitamin C (ascorbic acid), mushroom tyrosinase enzyme obtained from mushroom, l -3,4-dihydroxyphenylalanine (l -DOPA), potassium dihydrogen orthophosphate (pH 6.5), potassium hydroxide, hydroquinone monomethyl ether, elastase enzyme, substrate: (N-methoxy-succinyl-Ala-Ala-Pro-Val-P-nitro-anilide), tris(hydroxymethyl)-methyl-2-aminoethane sulfonate (TES) buffer, HEPES buffer which is abbreviation for (4-2-hydroxyethyl-l -piperazine ethane sulfonic acid), pH = 7.5, elafin positive control, collagenase type 1 from Clostridium histolyticum, FALGPA substrate (N-[3-(2-furyl)acryloyl]-Leu-Gly-Pro-Ala) and epigallocatechin gallate (EGCG) were purchased from Sigma Aldrich. UV recordings were made on ELX 808 (Bio Tek Instrumental, Italy), plates were purchased from (Mekkawy, Egypt).
Mushroom tyrosinase enzyme
Manufactured by Merck Group
Sourced in United States
Mushroom tyrosinase enzyme is a naturally occurring enzyme derived from mushrooms. Its core function is to catalyze the conversion of tyrosine to melanin, a pigment responsible for coloration in living organisms.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Promega (3 mentions)
Streptomycin, by Promega (3 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Thermo Fisher Scientific (3 mentions)
Muller hinton broth, by bioMérieux (3 mentions)
Griess reagent system, by Promega (3 mentions)
Dimethyl sulfoxide (dmso), by Promega (3 mentions)
P iodonitrotetrazolium chloride int, by Merck Group (2 mentions)
Tryptic soy broth (tsb), by bioMérieux (2 mentions)
Blood agar with 7 sheep blood, by bioMérieux (2 mentions)
Macconkey agar plates, by bioMérieux (2 mentions)
L tyrosine, by Merck Group (2 mentions)
N methoxysuccinyl ala ala pro val p nitroanilide, by Merck Group (1 mentions)
Epigallocatechin gallate (egcg), by Merck Group (1 mentions)
L 3 4 dihydroxyphenylalanine l dopa, by Merck Group (1 mentions)
Ascorbic acid vitamin c, by Merck Group (1 mentions)
Precision plus protein dual color standard, by Bio-Rad (1 mentions)
Sodium phosphate, by Avantor (1 mentions)
Coomassie brilliant blue g 250, by Avantor (1 mentions)
Dithiothreitol (dtt), by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
8 protocols using mushroom tyrosinase enzyme
1
Antioxidant and Anti-tyrosinase Assays
2
Purification and Conjugation of Biomolecules
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Unless stated otherwise,
all chemicals and solvents were obtained from Sigma-Aldrich or Fisher
Scientific and used as received. Dimethyl sulfoxide (DMSO) (molecular
biology grade) and mushroom tyrosinase enzyme was obtained from Sigma-Aldrich.
Dithiothreitol (DTT) was obtained from Fisher Scientific. 2-Mercaptoethanol
(BME) and tetramethylenediamine were obtained from Acros. Sodium dodecyl
sulfate (SDS), bromophenol blue, coomassie brilliant blue G250, sodium
phosphate, sodium chloride, and Tris were obtained from VWR. Precision
Plus Protein Dual Color Standards was obtained from Bio-Rad. Glycine
was obtained from Applichem. BCN-POE3-NH–lissamine rhodamine
B conjugate was obtained from SynAffix. MeTz-TAMRA was obtained from
BroadPharm. Trifluoroacetic acid (TFA), formic acid (FA), and MeCN
were obtained from BioSolve.
Trastuzumab antibodies were ordered
from Evitria AG and purified by Hitrap protein A HP (ProtA) column
(5 mL) on an Agilent 1260 Preparative HPLC with diode-array detector
(Section 4.6 ). The
antibody, present in sterile filtered CHO media samples, were applied
to the ProtA column without any pretreatment.
Sortase A,42 (link) UCHT1 variants,15 (link) and IL-2 variants14 (link),43 (link) were obtained
and purified as reported.
all chemicals and solvents were obtained from Sigma-Aldrich or Fisher
Scientific and used as received. Dimethyl sulfoxide (DMSO) (molecular
biology grade) and mushroom tyrosinase enzyme was obtained from Sigma-Aldrich.
Dithiothreitol (DTT) was obtained from Fisher Scientific. 2-Mercaptoethanol
(BME) and tetramethylenediamine were obtained from Acros. Sodium dodecyl
sulfate (SDS), bromophenol blue, coomassie brilliant blue G250, sodium
phosphate, sodium chloride, and Tris were obtained from VWR. Precision
Plus Protein Dual Color Standards was obtained from Bio-Rad. Glycine
was obtained from Applichem. BCN-POE3-NH–lissamine rhodamine
B conjugate was obtained from SynAffix. MeTz-TAMRA was obtained from
BroadPharm. Trifluoroacetic acid (TFA), formic acid (FA), and MeCN
were obtained from BioSolve.
Trastuzumab antibodies were ordered
from Evitria AG and purified by Hitrap protein A HP (ProtA) column
(5 mL) on an Agilent 1260 Preparative HPLC with diode-array detector
(
antibody, present in sterile filtered CHO media samples, were applied
to the ProtA column without any pretreatment.
Sortase A,42 (link) UCHT1 variants,15 (link) and IL-2 variants14 (link),43 (link) were obtained
and purified as reported.
3
Antioxidant and Tyrosinase Inhibitory Activity of Hemp Seed
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The hemp seed was purchased from “Nature
and Life” company in Hongcheon in South Korea and dried under
ambient conditions, before use. Reagents, such as ABTS, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide (MTT), α-MSH, Folin-Ciocalteu reagent, and mushroom
tyrosinase enzyme, were obtained from Sigma-Aldrich (St. Louis, MO,
USA). Potassium persulfate (K2S2O8) and 3,4-dihydroxy-l -phenylalanine (L-DOPA) were purchased
from Alfa Aesar (Haverhill, MA, USA).
and Life” company in Hongcheon in South Korea and dried under
ambient conditions, before use. Reagents, such as ABTS, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide (MTT), α-MSH, Folin-Ciocalteu reagent, and mushroom
tyrosinase enzyme, were obtained from Sigma-Aldrich (St. Louis, MO,
USA). Potassium persulfate (K2S2O8) and 3,4-dihydroxy-
from Alfa Aesar (Haverhill, MA, USA).
4
Phytochemical and Bioactivity Characterization
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Acetonitrile 99.9% was of high-performance liquid chromatography (HPLC) grade from Lab-Scan (Lisbon, Portugal), methanol was of analytical grade and supplied by Pronalab (Lisbon, Portugal). 2,2-Diphenyl-1-picrylhydrazyl (DPPH) was obtained from Alfa Aesar (Ward Hill, MA, USA). Dulbecco’s modified Eagle’s minimum essential medium (DMEM), fetal bovine serum (FBS), penicillin, streptomycin, Griess reagent system (Promega), DMSO, sulphorodamine B (SRB), lipopolysaccharide (LPS), 3,4-dihydroxy-l -phenylalanine (l -DOPA), and mushroom tyrosinase enzyme were obtained from Sigma-Aldrich Co. (Saint Louis, MO, USA). The culture media Muller Hinton broth (MHB) and Tryptic Soy Broth (TSB) were obtained from Biomerieux (Marcy l’Etoile, France). Blood agar with 7% sheep blood and Mac Conkey agar plates were purchased from Biomerieux Marcy l’Etoile, France). The dye p-iodonitrotetrazolium chloride (INT) was purchased from Sigma-Aldrich (Spruce Street; St. Louis, MO, USA) and was used as microbial growth indicator.
5
Anti-tyrosinase Activity Assay of Edible Bird's Nest
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The assay was based on the formation of L-dopachrome by the action of mushroom tyrosinase enzyme on its substrate, l -tyrosine (Sigma-Aldrich, Germany) [35 (link)]. An aliquot of 10 μL of tyrosinase (250 U/mL, Sigma-Aldrich, Germany) was supplemented to the wells of a 96-well plate containing 60 μL of diluted unhydrolyzed EBN, HPJ, HPP, HAJ, HBM, and HCJ from the hydrolysis at 0 and 4 h at the initial EBN concentrations of 0.0325, 0.075, 0.15, 0.3, and 0.6 mg/mL. This step was followed by the incubation of the plate at room temperature for 20 min and the addition of 140 μL of l -tyrosine (0.3 mg/mL) in phosphate buffer (pH 6.8). The absorbance of L-dopachrome at 480 nm was monitored in a microplate reader (VICTOR Nivo Filter, PerkinElmer, USA). The anti-tyrosinase ability of the samples was calculated as follows: Where.
B1: Absorbance of
B2: Absorbance of the sample after reacting with tyrosinase
B3 and B4: Absorbance of the blank for the negative control and sample, respectively.
The IC50 value of sample for the anti-tyrosinase capacity was calculated by GraphPad Prism software (version 5.0, Insightful Science LLC, USA).
6
Phytochemical Screening and Bioactivity Assays
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Acetonitrile 99.9% was of high-performance liquid chromatography (HPLC) grade, supplied by Lab-Scan (Lisbon, Portugal), and methanol was of analytical grade from Pronalab (Lisbon, Portugal). 2,2-Diphenyl-1-picrylhydrazyl (DPPH) was obtained from Alfa Aesar (Ward Hill, MA, USA). Dulbecco’s modified Eagle’s minimum essential medium (DMEM), fetal bovine serum (FBS), penicillin, streptomycin, Griess reagent system (Promega), dimethyl sulfoxide (DMSO), sulforhodamine B (SRB), lipopolysaccharide (LPS), 3,4-dihydroxy-L-phenylalanine (L-DOPA), dexamethasone, and mushroom tyrosinase enzyme were obtained from Sigma-Aldrich Co. (Saint Louis, MO, USA). Muller Hinton broth (MHB) and Tryptic Soy Broth (TSB) medium were purchased from Biomerieux (Marcy l’Etoile, France). Blood agar with 7% sheep blood and Mac Conkey agar plates were purchased from Biomerieux (Marcy l’Etoile, France). The p-iodonitrotetrazolium chloride (INT) dye and phenolic compound standards were purchased from Sigma-Aldrich (St. Louis, MO, USA).
7
Tyrosinase Inhibition Assay with Melanogenic Compounds
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l -Tyrosine, l -3,4-dihydroxyphenylalanine (l -DOPA), arbutin, mushroom tyrosinase enzyme (EC 1.14.18.1), and α-melanocyte-stimulating hormone (α-MSH) were obtained from Sigma Aldrich (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), and penicillin-streptomycin were purchased from Gibco-BRL Life Technologies (Grand Island, NY, USA). Primary tyrosinase (TYR) antibodies, β-actin, and horseradish peroxidase-conjugated secondary antibodies were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). Dipotassium phosphate and monopotassium phosphate were obtained from Junsei Chemical Co. Ltd. (Tokyo, Japan) and Yakuri Pure Chemicals Co. Ltd. (Osaka, Japan), respectively. All other reagents and solvents were purchased from E. Merck, Fluka, and Sigma-Aldrich unless otherwise stated.
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8
Reagent Standardization for Analytical Studies
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Acetonitrile 99.9% was of high-performance liquid chromatography (HPLC) grade from Lab-Scan (Lisbon, Portugal), methanol was of analytical grade and supplied by Pronalab (Lisbon, Portugal). 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) was obtained from Alfa Aesar (Ward Hill, MA, USA). Dulbecco's modified Eagle's minimum essential medium (DMEM), p-hydroxybenzoic, p-coumaric, protocatechuic and cinnamic acids, fetal bovine serum (FBS), penicillin, streptomycin, Griess reagent system (Promega), DMSO, sulphorodamine B (SRB), lipopolysaccharide (LPS), 3,4-dihydroxy-L-phenylalanine (L-DOPA) and mushroom tyrosinase enzyme were obtained from Sigma-Aldrich Co. (Saint Louis, MO, USA). Sodium alginate was provided by Fluka Chemie (USA), calcium chloride and Tween 80 by Panreac (Spain). Muller Hinton broth (MHB) and Tryptic Soy Broth (TSB) culture media were obtained from Biomerieux (Marcy l'Etoile, France). Blood agar (7% sheep blood), and MacConkey agar plates were purchased from Biomerieux Marcy l'Etoile, France). The p-iodonitrotetrazolium chloride (INT) was purchased from Sigma-Aldrich (Spruce Street; St. Louis, MO, USA) being used as microbial growth indicator.
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