Monomethoxy-capped polyethylene glycol (mPEG2k; Mw = 2000 g mol−1) was purchased from Sigma Aldrich and azeotropically distilled using dried toluene. Ammonium persulphate (APS), dimethylaminopyridine (DMAP), triethylamine (ET3N), and 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydrogen peroxide (H2O2), double-stranded salmon testes sperm DNA (SS-DNA), tris borate EDTA (TBE) buffer, trichloroacetic acid (TCA), and agarose of analytical grade were purchased from Sigma Aldrich. Trifluoroacetic acid (TFA) dimethylsulfoxide (DMSO), and 4-toluenesulfonyl chloride of analytical grade were purchased from Merk Millipore. Dried tetrahydrofuran (THF), dichloromethane (DCM), ethyl acetate, and diethyl ether were obtained from an Anhydrous Engineering dry solvent system based on Grubbs design25 (link).
Trichloroacetic acid tca
Manufactured by Merck Group
Sourced in United States, Germany, India, Italy, United Kingdom, China, Sao Tome and Principe, Macao, Australia
Trichloroacetic acid (TCA) is a chemical compound commonly used in analytical and research applications. It is a colorless, crystalline solid that is soluble in water and organic solvents. TCA is a strong acid and is often used as a precipitating agent for proteins and other biomolecules in various laboratory procedures.
Automatically generated - may contain errors
Lab products found in correlation
Thiobarbituric acid tba, by Merck Group (45 mentions)
Gallic acid, by Merck Group (36 mentions)
Acetic acid, by Merck Group (32 mentions)
Bovine serum albumin (bsa), by Merck Group (26 mentions)
Methanol, by Merck Group (25 mentions)
Thiobarbituric acid, by Merck Group (23 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (23 mentions)
Sulforhodamine b srb, by Merck Group (21 mentions)
Ascorbic acid, by Merck Group (21 mentions)
Sodium hydroxide (naoh), by Merck Group (21 mentions)
Hydrochloric acid (hcl), by Merck Group (21 mentions)
Quercetin, by Merck Group (18 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (18 mentions)
Trypan blue, by Merck Group (16 mentions)
Folin ciocalteu reagent, by Merck Group (16 mentions)
Ellipticine, by Merck Group (15 mentions)
Ethanol, by Merck Group (15 mentions)
Acetonitrile, by Merck Group (14 mentions)
Sodium acetate, by Merck Group (14 mentions)
Sodium carbonate, by Merck Group (14 mentions)
355 protocols using trichloroacetic acid tca
1
Synthesis of Functionalized mPEG-Based Materials
2
Cytotoxicity Assessment of Secoisolariciresinol Diglucoside
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RPMI-1640 Medium, fetal bovine serum, dimethylsulfoxide (DMSO), Ellman’s reagent [5,5-Dithio-bis-(2-nitro bezoic acid)], β-mercaptoethanol, glutathione (GSH), glutathione reductase, sodium dodecyl sulfate (SDS), sodium bicarbonate, 1,1.3,3-tetramethoxypropane, trichloroacetic acid (TCA) and thiobarbituric acid were all purchased from Sigma Aldrich Chemical Co. (St. Louis, MO, USA). Secoisolariciresinol diglucoside (SDG) for HPLC analysis was purchased from ChromaDex (Santa Ana, CA, USA). Triton X-100, penicillin, streptomycin were procured from MP Biochemical (Santa Ana, California, USA). All other chemical reagents and extraction solvents were of analytical grade, and all analysis solvents were of HPLC grade and they were obtained from E-Merck. Tamoxifen (TAM) was obtained from Sigma Aldrich Chemical Co. (St. Louis, MO, USA). Each vial of TAM contains one gm white powder.
3
Measuring Protein Synthesis by Radiolabeled Leucine
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Protein synthesis was measured by incorporation of Leucine L-(4,5-3H) (obtained from Perkin Elmer (NET1166005MC, Waltham, MA, USA). Cells were treated for drugs as indicated for 5 h and washed with PBS and RPMI-1640 media and 3.5 μCi/mL Leucine L-(4,5-3H) was added for 1 h. Cell suspensions were placed on Whatman GF/C microfiber filters and proteins were precipitated with ice-cold 5% trichloroacetic acid (TCA) (Sigma-Aldrich, St Louis, MO, USA). Filters were washed twice with 5% TCA, once with 99% ethanol, and left to dry. Radioactivity was measured using liquid scintillation counting.
4
Hyaluronidase-Mediated Synthesis of Oligosaccharides
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o-HA (6~10 disaccharides, 4-5 kDa) was prepared by hyaluronidase degradation methods 29 (link), 30 (link). One hundred milligram of h-HA was dissolved in 50 mL of 0.1 M sodium acetate buffer (pH 5.4) containing 0.15 M NaCl, and then digested by 20000 units of bovine testicular hyaluronidase (Sigma) incubating at 37 oC. At regular intervals (2, 4, 6, 8, and 24 h), 10 mL aliquots were removed and the reaction was terminated by the addition of 1 mL of 100% Trichloroacetic acid (TCA, Sigma). The precipitate was eliminated by centrifugation and the supernatants from different time was mixed. The aliquots were dialyzed against distilled water with dialysis membrane (MWCO, 1000 Da) for 72 h at 4 oC. The internal solution was dialyzed against distilled water with dialysis membrane (MWCO, 5000 Da) for another 72 h. At last, the external phase was collected, followed by lyophilized. Lyophilized o-HA was preserved at 4 oC.
The o-HA samples were further characterized by Gel Permeation Chromatography (GPC) to determine macromolecular weight and weight distribution. The samples were dissolved in chloroform and cast on KBr plate, and the Fourier Transforms Infrared Spectroscopy (FTIR) spectra were kept. Nuclear Magnetic Resonance Analysis (1H NMR) was used to characterize chemical composition.
The o-HA samples were further characterized by Gel Permeation Chromatography (GPC) to determine macromolecular weight and weight distribution. The samples were dissolved in chloroform and cast on KBr plate, and the Fourier Transforms Infrared Spectroscopy (FTIR) spectra were kept. Nuclear Magnetic Resonance Analysis (1H NMR) was used to characterize chemical composition.
5
Oxidative Stress and Inflammatory Biomarkers Assay
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CC14, Tris-HCl, thiobarbituric acid (TBA), oxidized and reduced glutathione, reduced β-nicotinamide adenine dinucleotide phosphate (NADPH), glucose-6-phosphate, 1-chloro-2,4-dinitrobenzene (CDNB), glutathione reductase, 5,5-dithio-bis-2-nitrobenzoic acid (DTNB), 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), potassium persulfate, sulfosalicylic acid (SSA), bovine serum albumin (BSA), hydrogen peroxide (H2O2), flavin adenine dinucleotide (FAD), 2,6-dichloroindophenol, trichloroacetic acid (TCA), Tween 20, and ethylenediaminetetraacetic acid (EDTA) were obtained from Sigma Chemical Co. (St. Louis, MO, USA). Rabbit polyclonal antibody specific for 8-hydroxy-2′-deoxyguanosine (8-8-OHdG), Rabbit polyclonal antibody specific for 4-hydroxy-2-nonenal (HNE), Rabbit polyclonal antibody specific to tumor necrosis factor alpha (TNF-α), Rabbit polyclonal antibody specific to interleukin 6 (IL-6), Rabbit polyclonal antibody specific to prostaglandin E2 (PGE2), EnVision™ + System/horseradish peroxidase (HRP), Rb (DAB+), target retrieval solution, and antibody diluent were purchased from Dako (Agilent Technologies Company, Denmark).
6
Quantitative Analysis of Biomarkers
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Acetonitrile (ACN), dichloromethane (DCM), di‐isopropyl ether (DIPE), ethanol (EtOH), ethyl acetate, formic acid, hexane and methanol (MeOH) were purchased from Merck (Darmstadt, Germany). Trichloroacetic acid (TCA) was obtained from Sigma Aldrich (Castle Hill, NSW, Australia). Water used was ultrapure grade from a Thermo Scientific Barnstead Smart2Pure water purification System (ThermoScientific; Langenselbold, Hungary). All solvents used were of HPLC grade.
Certified reference standards for HC and C were obtained from Sigma Aldrich (Castle Hill, NSW, Australia). Internal standard D4‐HC was obtained from Cambridge Isotope Laboratories (Andover, Massachusetts, USA). TACA was obtained from Toronto Research Chemicals (Toronto, Ontario, Canada).
Certified reference standards for HC and C were obtained from Sigma Aldrich (Castle Hill, NSW, Australia). Internal standard D4‐HC was obtained from Cambridge Isotope Laboratories (Andover, Massachusetts, USA). TACA was obtained from Toronto Research Chemicals (Toronto, Ontario, Canada).
7
Antioxidant Evaluation of Plant Extracts
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Ascorbic acid; aluminum chloride, ferric chloride (FeCl3); Folin-Ciocalteu; bovine serum albumin (BSA); potassium persulphate; 2,2-diphenyl-1-picrylhy-drazyl (DPPH); nitro blue tetrazolium (NBT); phenazine methosulphate (PMS); sulphosalicylic acid; thiobarbituric acid (TBA) and trichloroacetic acid (TCA) were purchased from Sigma chemicals (St. Louis, MO, USA). All other chemicals procured from Sysco Research Laboratory (Mumbai, India). Methanol (99.8%) used were of analytical grade and purchased from Merck Life Science Private Limited, Mumbai, India.
8
Comprehensive Analytical Protocol for Plant Composition
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Methanol (MeOH) and chloroform HPLC grade were purchased from Fluka Chemika (Steinheim, Switzerland). Sodium carbonate (Na2CO3), sodium phosphate, sodium tetraborate decahydrate, sodium hydroxide (NaOH), Kjeldahl catalyst, ethanol, acetone, phenol, acid boric, potassium acetate (KCH3CO2), potassium hydroxide, and standard glucose were from E. Merck (Darmstadt, Germany). Derivatization reagent (14% boron trifluoride in methanol) was purchased from Alltech Associates (Deerfield, IL, USA). Total Dietary Fiber Assay Kit (TDF-100A Kit) that contains α-amylase (A3306), amyloglucosidase (A9913), protease (P3910) and celite, fatty acids, methyl esters standards “FAME Mix C4-C24 (18919 Supelco)” and “PUFA No. 1 Marine Source (47033 Supelco)”, potassium ferricyanide, iron (II) chloride, iron (III) chloride, ferrozine, trichloroacetic acid (TCA), Folin–Ciocalteu reagent (FCR), phloroglucinol, rutin, butylated hydroxytoluene (BHT), ethylenediaminetetraacetic acid (EDTA), 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydrochloric acid (HCl), aluminum chloride (AlCl3), sulphuric acid, hexane, and petroleum ether were obtained from Sigma-Aldrich (St. Louis, MO, USA).
9
Bacterial Cultivation and Protein Extraction Protocol
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Bacterial cultivation and extracellular protein extraction were
carried out as described previously.19 (link) Briefly,
all bacterial isolates were grown in triplicate overnight (14–16
h) in 10 mL tryptic soy broth (TSB, OXOID, Basingstoke, UK) under
vigorous shaking (115 rpm) at 37 °C in a water bath. The cultures
were then diluted into 10 mL prewarmed Roswell Park Memorial Institute
1640 (RPMI) medium supplemented with 2 mM glutamine (GE Healthcare/PAA,
Little Chalfont, United Kingdom) to an optical density at 600 nm (OD600) of 0.1 and cultivation was continued under the same conditions.
Exponentially growing cells with an OD600 of ±0.5
were again diluted into 20 mL of fresh prewarmed RPMI 1640 medium
to a final OD600 of 0.1 and their cultivation was continued
until an OD600 of ±1.3 was reached, which corresponds
to the stationary growth phase. Then, growth medium fractions were
collected by centrifugation. Proteins in the growth medium were precipitated
overnight with 10% trichloroacetic acid (TCA, Sigma-Aldrich, St. Louis,
USA) on ice. The precipitated proteins were collected by centrifugation.
Pellets of precipitated proteins were washed once with ice-cold acetone,
dried at room temperature and stored at −20 °C until further
use.
carried out as described previously.19 (link) Briefly,
all bacterial isolates were grown in triplicate overnight (14–16
h) in 10 mL tryptic soy broth (TSB, OXOID, Basingstoke, UK) under
vigorous shaking (115 rpm) at 37 °C in a water bath. The cultures
were then diluted into 10 mL prewarmed Roswell Park Memorial Institute
1640 (RPMI) medium supplemented with 2 mM glutamine (GE Healthcare/PAA,
Little Chalfont, United Kingdom) to an optical density at 600 nm (OD600) of 0.1 and cultivation was continued under the same conditions.
Exponentially growing cells with an OD600 of ±0.5
were again diluted into 20 mL of fresh prewarmed RPMI 1640 medium
to a final OD600 of 0.1 and their cultivation was continued
until an OD600 of ±1.3 was reached, which corresponds
to the stationary growth phase. Then, growth medium fractions were
collected by centrifugation. Proteins in the growth medium were precipitated
overnight with 10% trichloroacetic acid (TCA, Sigma-Aldrich, St. Louis,
USA) on ice. The precipitated proteins were collected by centrifugation.
Pellets of precipitated proteins were washed once with ice-cold acetone,
dried at room temperature and stored at −20 °C until further
use.
10
Analytical Reagents for Glutathione Assay
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All chemicals and reagents used in this study were of analytical purity, being purchased from different providers: acetonitrile was purchased from VWR International, SAS, Fontenay-sous-Bois, France, anhydrous disodium phosphate (Na2HPO4), and 85% phosphoric acid solution (H3PO4) were purchased from Merck KGaA (Darmstadt, Germany). The Ellman’s reagent, trichloroacetic acid (TCA), ethylenediaminetetraacetate (EDTA-Na2) powder, sodium chloride (NaCl) powder, phosphate buffer solution (PBS), GSH and GSSG powders were all purchased from Sigma-Aldrich (Darmstadt, Germany). Ultra pure water was obtained using the Milli-Q purification system (Merck Millipore Corporation, Burlington, MA, USA).
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