Plant-derived natural products and synthetic molecules with known or expected anti-inflammatory activity were purchased from different vendors. HA15 ((N-[4-[3-[[5-(dimethyl amino) naphthalen-1-yl]sulfonyl amino]phenyl]-1,3-thiazol-2-yl] acetamide, Calbiochem) was purchased from Sigma-Aldrich (St. Luis, MS). Geranylgeranyl acetone (GGA) and sodium salt of phenyl butyrate (PBA) were from Selleck Chemicals GmbH (Berlin, Germany). 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) and phenazine methosulfate (PMS) were from Thermo Fisher Scientific (Waltham, MA, USA) or (TCI, Tokyo, Japan), respectively. OxPAPC was prepared from 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (Avanti Polar Lipids, Birmingham, AL, USA) and analysed as described (Watson et al. 1997 (link)). OxPAPC was stored as a chloroform solution at − 80 °C, and evaporated under the stream of argon before solubilisation in the cell culture medium.
2 3 bis 2 methoxy 4 nitro 5 sulfophenyl 2h tetrazolium 5 carboxanilide xtt
Manufactured by Thermo Fisher Scientific
Sourced in United States
XTT is a water-soluble tetrazolium salt that is used as a colorimetric assay for the measurement of cell viability and proliferation. When XTT is reduced by viable cells, it forms a colored formazan product that can be quantified spectrophotometrically.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Phenazine methosulfate, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Osmium tetroxide, by Electron Microscopy Sciences (2 mentions)
Sodium chloride (nacl), by Thermo Fisher Scientific (2 mentions)
Tween 20, by Thermo Fisher Scientific (2 mentions)
1 palmitoyl 2 arachidonoyl sn glycero 3 phosphocholine, by Avanti Polar Lipids (1 mentions)
Human recombinant insulin, by Thermo Fisher Scientific (1 mentions)
Cisplatin, by Bio-Techne (1 mentions)
Bromophenol blue, by Thermo Fisher Scientific (1 mentions)
Salicylaldehyde, by Merck Group (1 mentions)
D l valine, by Merck Group (1 mentions)
O vanillin, by Merck Group (1 mentions)
Vivoso4 3h2o, by Merck Group (1 mentions)
3 5 dinitrosalicylic acid, by Merck Group (1 mentions)
α amylase from porcine pancreas, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
Glycerol, by Merck Group (1 mentions)
6 protocols using 2 3 bis 2 methoxy 4 nitro 5 sulfophenyl 2h tetrazolium 5 carboxanilide xtt
1
Anti-inflammatory natural product evaluation
2
Evaluation of α-Amylase Inhibitory Activity
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The reagents used in this study were analytically pure and were purchased from commercial manufacturers. α-Amylase from porcine pancreas, bovine serum albumin (BSA), 3, 5-dinitrosalicylic acid (DNS), D-/L-valine, dimethyl sulfoxide (DMSO), Dulbecco’s modified Eagle’s medium (DMEM), 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES), ethylenediaminetetraacetic acid (EDTA) tetrasodium salt dihydrate, glycerol, 2-mercaptoethanol, salicylaldehyde, sodium orthovanadate (Na3VVO4), sodium chloride, sodium hydroxide, sodium dodecyl sulfate (SDS), p-Nitrophenyl Phosphate (pNPP), starch, o-vanillin, Tris, VIVOSO4•3H2O were from SIGMA-Aldrich (Merck KGaA, Darmstadt, Germany). Acarbose, bromophenol blue, fetal bovine serum, human recombinant insulin, penicillin, streptomycin, 2, 3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT), and phenazine methosulfate were from Thermo Fisher Scientific (Waltham, MA, USA). Cisplatin was from Tocris Bioscience (Minneapolis, MN, USA). The 96-well UV/ Vis micro test plates and cell culture dishes were from Corning (New York, NY, USA)/ Ratiolab GmbH (Dreieich, Germany) and TPP® (Trasadingen, Switzerland).
3
Synthesis and Characterization of Thermoresponsive Nanocarriers
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N-isopropylacrylamide (NIPAAm), purchased from Aldrich Chemical Corp. (Milwaukee, WI, USA), was recrystallized with hexane. 1-Vinylimidazole (VI), N,N′-azobisisobutyronitrile (AIBN), 1,4-dioxane, and cyclohexane were supplied from Fluka AG (Buchs, Switzerland). Soybean lecithin was obtained from Iassyfarm S.A. (Iassy, Romania). Doxorubicin hydrochloride (DXR) was supplied from Sigma-Aldrich (St. Louis, MO, USA)
Reagents and consumables used to perform biological assays were purchased from different sources as follows: Dulbecco’s modified Eagle’s medium (DMEM), paraformaldehyde (PFA), and Triton X-100, were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany); 2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide (XTT), phenazine methosulfate (PMS), 4′,6-diamidino-2-phenylindole (DAPI), fetal bovine serum, and antibiotics were from Thermo Fisher Scientific (Waltham, MA, USA); cell culture dishes were from TPP® (Trasadingen, Switzerland), black 96-well micro test plates, F-bottom were from Ratiolab (Ratiolab GmbH, Dreieich, Germany); phalloidin-Fluorescein isothiocyanate (FITC) was from Tocris Bioscience (Bio-Techne Ltd., Minneapolis, MN, USA).
Reagents and consumables used to perform biological assays were purchased from different sources as follows: Dulbecco’s modified Eagle’s medium (DMEM), paraformaldehyde (PFA), and Triton X-100, were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany); 2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide (XTT), phenazine methosulfate (PMS), 4′,6-diamidino-2-phenylindole (DAPI), fetal bovine serum, and antibiotics were from Thermo Fisher Scientific (Waltham, MA, USA); cell culture dishes were from TPP® (Trasadingen, Switzerland), black 96-well micro test plates, F-bottom were from Ratiolab (Ratiolab GmbH, Dreieich, Germany); phalloidin-Fluorescein isothiocyanate (FITC) was from Tocris Bioscience (Bio-Techne Ltd., Minneapolis, MN, USA).
4
Fabrication and Characterization of Polyelectrolyte Multilayers
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Catheter tubes (polyethylene, PE-100, inner
diameter =0.034 in.) were obtained from Intramedic, Becton Dickinson
& Co. Poly-l -lysine hydrobromide (PLL, MW = 15,000–30,000),
poly-l -glutamic acid Na salt (PGA, MW = 50 000–100 000),
chitosan (CH, medium molecular weight) and branched polyethylenimine
(BPEI, MW = 25 000) were purchased from Sigma-Aldrich. Sodium
hyaluronate (HA, MW = 1 500 000–2 200 000)
was purchased from Acros Organic. 2,3-Bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide
(XTT) and RPMI 1640 powder containingl -glutamine and phenol
red (without HEPES or Na bicarbonate) were obtained from Invitrogen.
Phosphate-buffered saline (PBS) concentrate was obtained from OmniPur,
and formaldehyde, glutaraldehyde, and menadione were obtained from
Sigma. Tween-20 was obtained from Acros, and osmium tetroxide (4%)
was obtained from Electron Microscopy Sciences. NaCl, Tris-base, and
3-(N-morpholino)propanesulfonic acid (MOPS) were obtained from Fisher
Scientific. All materials were used as received.
diameter =0.034 in.) were obtained from Intramedic, Becton Dickinson
& Co. Poly-
poly-
chitosan (CH, medium molecular weight) and branched polyethylenimine
(BPEI, MW = 25 000) were purchased from Sigma-Aldrich. Sodium
hyaluronate (HA, MW = 1 500 000–2 200 000)
was purchased from Acros Organic. 2,3-Bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide
(XTT) and RPMI 1640 powder containing
red (without HEPES or Na bicarbonate) were obtained from Invitrogen.
Phosphate-buffered saline (PBS) concentrate was obtained from OmniPur,
and formaldehyde, glutaraldehyde, and menadione were obtained from
Sigma. Tween-20 was obtained from Acros, and osmium tetroxide (4%)
was obtained from Electron Microscopy Sciences. NaCl, Tris-base, and
3-(N-morpholino)propanesulfonic acid (MOPS) were obtained from Fisher
Scientific. All materials were used as received.
5
Cytotoxicity Assessment of Milk Fractions
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IPEC-J2 and Caco-2/15 cells were seeded at a density of 104 cells/well in a ninety-six-well plate and allowed to adhere to the plate overnight. Cells were further incubated for 24 h with increasing doses of milk fractions (0, 0·1, 1 and 10 mg/ml) in the serum-deprived culture medium. Cell proliferation was measured using the 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT; InVitrogen), which assess cell viability and proliferation as a function of redox potential. Briefly, fresh 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide stock solution (1 mg/ml in PBS) was prepared, and PMS (phenazine methosulfate 15 mg/ml in PBS, store in dark and −20°C) was diluted 1:100 in PBS. PMS was added to 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide solution (40:1), and 50 µl of the mix was added to each wells. Cells were incubated for 1 h before the absorbance was measured at 450/630 nm. The absorbance in untreated cells (0·0 mg/ml) was set as 100 %. In total, four independent experiments were done in triplicate.
6
Polyelectrolyte Coatings for Cell Adhesion
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