Sucrose derived from sugarcane was supplied by C&H (ASR Group, West Palm Beach, FL, USA) which was used as the primary carbon source for the production of vanillin by fermentation in the engineered yeast strain. Inorganic salts and trace metals were Reagent Plus grade (Sigma-Aldrich, St. Louis, MO, USA). Vitamins, L-lysine HCl, maltose, myo-inositol and succinate were Bioreagent grade (Sigma-Aldrich, St. Louis, MO, USA). Ethyl acetate and hexanes were reagent grade (Acros Organics, Fair Lawn, NJ, USA). Water and acetonitrile were HPLC grade (Acros Organics, Fair Lawn, NJ, USA). Silica gel was ultrapure grade, 60-200μ (Acros Organics, Fair Lawn, NJ, USA).
Bioreagent grade
Manufactured by Merck Group
Sourced in United States
BioReagent grade is a high-quality laboratory reagent designed for use in biological and biochemical research applications. It is manufactured to meet stringent purity and consistency standards, ensuring reliable and reproducible results. The core function of BioReagent grade is to provide a reliable and standardized source of reagents for scientists working in life science and biotechnology fields.
Automatically generated - may contain errors
Lab products found in correlation
Reagentplus, by Merck Group (2 mentions)
Hplc grade, by Thermo Fisher Scientific (1 mentions)
Silica gel, by Thermo Fisher Scientific (1 mentions)
Na2haso4 7h2o, by Merck Group (1 mentions)
Cm 0602, by Procell (1 mentions)
Pm150312, by Procell (1 mentions)
Pb180120, by Procell (1 mentions)
Nextera xt dna library preparation kit, by Illumina (1 mentions)
Trypsin gold, by Promega (1 mentions)
Iodoacetamide, by Merck Group (1 mentions)
Lc ms grade formic acid, by Thermo Fisher Scientific (1 mentions)
Lc ms grade acetonitrile, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Fluoxetine hydrochloride, by Merck Group (1 mentions)
Tricaine methanesulfonate, by Merck Group (1 mentions)
Milli q system, by Thermo Fisher Scientific (1 mentions)
Sertraline hydrochloride, by Merck Group (1 mentions)
Paroxetine hydrochloride hemihydrate, by Merck Group (1 mentions)
7 protocols using bioreagent grade
1
Vanillin Production by Engineered Yeast
2
Preparation of Arsenic Stock Solutions
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Distilled water from Tecnología y Control Ambiental (>1.0 MΩ-cm) was used to prepare all solutions and dilute samples. A solution of 1 M HCl was prepared from concentrated hydrochloric acid (38%, Ecolaboratorios/Karal, ACS reagent grade, CAS: 7647-01-0). A solution of 0.5 M H2SO4 was prepared from concentrated sulphuric acid (98%, Ecolaboratorios/Karal, ACS reagent grade, CAS: 7664-93-9). A stock solution of As(V) (1000 mg L-1) was prepared by dissolving sodium arsenate dibasic hydrate (Na2HAsO4.7H2O, Sigma, ≥98.0% purity, CAS: 10048-95-0) in distilled water, with the pH adjusted to 7.5 with the addition of a small volume of 1 M HCl. A stock solution of As(III) (1000 mg L-1) was prepared by dissolving arsenic trioxide (As2O3, Sigma-Aldrich, ReagentPlus®, ≥99.0% purity, CAS: 1327-53-3) in a small volume of 1.0 N NaOH (Sigma, BioReagent grade, CAS: 1310-73-2) and making up to the desired volume with distilled water. The stock solution was adjusted to pH 5 with the addition of a small volume of 1 M HCl. Arsenic standard solutions (1000 μg L-1) were prepared from the 1000 mg L-1 stock solutions by diluting with distilled water. All arsenic standard solutions were stored in the dark at 3°C to prevent changes in the speciation.
3
Mouse Liver Cell Line Cultivation
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Alpha mouse liver 12 (AML-12) cell line was established from mouse hepatocytes (source: a 3-month-old CD1 strain MT42 mouse line) that ectopically express human transforming growth factor alpha (ATCC #CRL-2254). AML12 cells were authenticated by their general morphology, short tandem repeat microsatellite marker analysis, capacity to form lipid droplets upon oleic acid treatment and RT-qPCR for mouse genes. AML-12 cells were cultured at 37 °C and 5% CO2 in DMEM/F12 (Procell #PM150312) supplemented with 10% fetal bovine serum (Procell #164210-50), 10 µg/mL insulin, 5.5 µg/mL transferrin, 5 ng/mL selenium, 40 ng/mL dexamethasone and 1% penicillin/streptomycin (100 U/mL and 0.1 mg/mL, respectively; Procell #PB180120), which is formulated as the AML-12 culture medium (Procell #CM-0602). Cells were seeded to 6-well plates (NEST #703001) or 10-cm dish (NEST #704002). When reaching 80-90% confluence, cells were treated in DMEM (Gibco #11966025) containing various concentrations of glucose (BioReagent grade, Sigma #G7021) for 8 h.
4
Single-cell RNA-seq of A375 Cells
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A375 cells were harvested and divided into two pools on each sampling day. One pool was used for extraction of genomic DNA, and the other pool was methanol fixed for Drop-seq analysis. Methanol fixation was performed as described previously51 . Briefly, cells were centrifuged at 300 g for 5 min and resuspended in 80% methanol (BioReagent grade; Sigma-Aldrich) and 20% PBS (Gibco). The resuspended cells were incubated on ice for at least 15 min and then stored at −80 °C. On the day of the Drop-seq experiment, cells were recovered by centrifugation at 2000g for 5 min, washed once with PBS-0.01% BSA, and resuspended in PBS-0.01% BSA to a concentration of 100 cells/μl. Finally, Drop-seq was performed as described previously23 (link).
Droplets were collected into 50-ml conical tubes over a 15 min time period. After which, the droplets were broken, and the RNAs on beads were subjected to reverse transcription.
Aliquots of 2,000 beads were amplified in each tube using the following PCR steps: 95 °C for 3 min, then four cycles of 98 °C for 20 s, 65 °C for 45 s, 72 °C for 3 min, and 11 cycles of 98 °C for 20 s, 67 °C for 20 s, 72 °C for 3 min, and 1 cycle of 72 °C for 5 min. The amplified products were purified with 0.6 × Ampure XP beads and fragmented, tagged, and amplified using a Nextera XT DNA Library Preparation kit (Illumina).
Droplets were collected into 50-ml conical tubes over a 15 min time period. After which, the droplets were broken, and the RNAs on beads were subjected to reverse transcription.
Aliquots of 2,000 beads were amplified in each tube using the following PCR steps: 95 °C for 3 min, then four cycles of 98 °C for 20 s, 65 °C for 45 s, 72 °C for 3 min, and 11 cycles of 98 °C for 20 s, 67 °C for 20 s, 72 °C for 3 min, and 1 cycle of 72 °C for 5 min. The amplified products were purified with 0.6 × Ampure XP beads and fragmented, tagged, and amplified using a Nextera XT DNA Library Preparation kit (Illumina).
5
Zebrafish Brain Perfusion and Photoelectrochemistry
6
Preparation of Alkylated Protein Digests
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Pooled gel filtration fractions of
extracted cross-linked proteins in the 400 kDa to 1 to 2 MDa range
were concentrated to ∼10 mg protein/mL with 0.5 mL of Amicon
Ultra 10 kDa cut off centrifugal filters (Millipore). Prior to digestion,
cysteines were alkylated by the addition of a solution of 0.8 M iodoacetamide
(Sigma–Aldrich), followed by the addition of solutions of 1
M Tris-HCl pH 8.0 and 9.6 M urea (Bioreagent grade, Sigma–Aldrich)
to obtain final concentrations of 40 mM iodoacetamide, 0.1 M Tris
HCl and 6 M urea, respectively. Incubation was for 30 min at room
temperature in the dark. The solution was diluted six times by the
addition of 0.1 M Tris–HCl pH 8.0 and digested with trypsin
(Trypsin Gold, Promega, Madison, WI) overnight at 30 °C at a
1:50 (w/w) ratio of enzyme and substrate. Peptides were desalted on
C18 reversed-phase TT3 top tips (Glygen, Columbia, MD), eluted with
0.1% TFA in 50% acetonitrile, and dried in a vacuum centrifuge.
extracted cross-linked proteins in the 400 kDa to 1 to 2 MDa range
were concentrated to ∼10 mg protein/mL with 0.5 mL of Amicon
Ultra 10 kDa cut off centrifugal filters (Millipore). Prior to digestion,
cysteines were alkylated by the addition of a solution of 0.8 M iodoacetamide
(Sigma–Aldrich), followed by the addition of solutions of 1
M Tris-HCl pH 8.0 and 9.6 M urea (Bioreagent grade, Sigma–Aldrich)
to obtain final concentrations of 40 mM iodoacetamide, 0.1 M Tris
HCl and 6 M urea, respectively. Incubation was for 30 min at room
temperature in the dark. The solution was diluted six times by the
addition of 0.1 M Tris–HCl pH 8.0 and digested with trypsin
(Trypsin Gold, Promega, Madison, WI) overnight at 30 °C at a
1:50 (w/w) ratio of enzyme and substrate. Peptides were desalted on
C18 reversed-phase TT3 top tips (Glygen, Columbia, MD), eluted with
0.1% TFA in 50% acetonitrile, and dried in a vacuum centrifuge.
7
Quantification of Selective Serotonin Reuptake Inhibitors
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Fluoxetine hydrochloride (CAS# 56296-78-7), Paroxetine hydrochloride hemihydrate (CAS# 110429-35-1), Sertraline hydrochloride (CAS# 79559-97-0), dimethyl sulfoxide (CAS# 67-68-5), tricaine methanesulfonate (CAS# 886-86-2), and water (sterile filtered, Bioreagent grade) were purchased from Sigma-Aldrich Canada (Oakville, ON, Canada). Working stocks of Fluoxetine hydrochloride and Paroxetine hydrochloride hemihydrate were made by dissolving the powders in water at 10 and 5 mg/mL, respectively. Sertraline hydrochloride was dissolved in dimethyl sulfoxide (DMSO) at 10 mg/mL. These stocks were stored at 4°C. Methanol used in SSRI extractions was distilled-in-glass grade (Caledon Laboratories Ltd., Georgetown, ON, Canada). Mobile phases used in the liquid chromatography–high resolution mass spectrometry (LC-HRMS) were prepared with deionized water obtained from a Milli-Q system and LC-MS grade acetonitrile and LC-MS grade formic acid (Fisher Chemical, Ottawa, ON, Canada).
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