Unless otherwise stated, all chemicals, solvents, and reference compounds were obtained from Sigma-Aldrich® (Steinheim, Germany) or Carl Roth GmbH & Co. KG (Karlsruhe, Germany) with the highest purity available. β-myrcene (purity ≥ 95%), racemic linalool (purity ≥ 97%), (R)-(−)-linalool (purity ≥ 99%), and geraniol (purity ≥ 99%) were purchased from Sigma-Aldrich® (Steinheim, Germany). pEHISTEV [22 (link)] and pMS470 [23 (link)] vectors were employed for expression. Restriction enzymes were acquired from Thermo Scientific (St. Leon-Rot, Germany). Sterile water was purchased from Fresenius Kabi (Graz, Austria). For cloning and plasmid replication, E. coli Top10 F′ (F′[lacIq Tn10(tetR)] mcrA Δ(mrr-hsdRMS-mcrBC) ϕ80lacZΔM15 ΔlacX74 deoR nupG recA1 araD139 Δ(ara-leu)7697 galU galK rpsL(StrR) endA1 λ−) from Life technologies (Vienna, Austria) was used. Recombinant Ldi was expressed in E. coli BL21-CodonPlus(DE3)-RP (F-ompT hsdS(rB− mB−) dcm+ Tetrgal endA Hte [argU proL Camr]) (Life technologies, Vienna, Austria).
Sterile water
Manufactured by Fresenius
Sourced in Germany, Austria, Sweden
Sterile water is a laboratory equipment product that provides purified water free from microorganisms and impurities. Its core function is to serve as a high-purity solvent for various laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Restriction enzyme, by Thermo Fisher Scientific (2 mentions)
Falcon tube, by Thermo Fisher Scientific (2 mentions)
Methanol, by Merck Group (2 mentions)
Geraniol, by Merck Group (1 mentions)
Racemic linalool, by Merck Group (1 mentions)
β myrcene, by Merck Group (1 mentions)
R linalool, by Merck Group (1 mentions)
Prestoblue cell viability reagent, by Thermo Fisher Scientific (1 mentions)
Infinite m200, by Tecan (1 mentions)
Bacto yeast extract, by BD (1 mentions)
Bacto peptone, by BD (1 mentions)
Zeocin, by InvivoGen (1 mentions)
Standard laboratory reagents, by Merck Group (1 mentions)
Difco yeast nitrogen base w o amino acids, by BD (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Methyl methanesulfonate, by Merck Group (1 mentions)
Sodium pyruvate, by Merck Group (1 mentions)
Mitomycin c, by Merck Group (1 mentions)
Absolute ethanol, by Merck Group (1 mentions)
Sodium bicarbonate, by Merck Group (1 mentions)
9 protocols using sterile water
1
Recombinant Ldi expression in E. coli
2
Cell Viability Assay with Peptides
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A total of 1 × 105 macrophages or WI-38 cells per well were distributed into a 96-well plate in corresponding cell culture medium. The controls were the media control, heat-inactivated cells and a diluent control (sterile water; Fresenius Kabi, Bad Homburg, Germany). The cells were then stimulated with the peptides at different concentrations overnight. Twenty microliters of PrestoBlue Cell Viability Reagent (Life Technologies, Darmstadt, Germany) was added per well and incubated for 20 min at 37 °C and a 5% CO2 atmosphere. Fluorescence was measured at 560 nm (excitation) and 600 nm (emission) with a TECAN infinite M200 microplate reader. The OD of the media control was then subtracted from the other results. The unstimulated cells were set to 100% viability [30 (link)].
3
Sputum Induction for Respiratory Research
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The sputum induction21 (link) was performed with an eFlow rapid nebulizer (PARI, Hamburg, Germany) filled with a hypertonic saline solution. The solution was prepared by mixing sterile water (Fresenius Kabi, Stockholm, Sweden) with 4% sodium chloride (B. Braun Medical AB, Stockholm, Sweden). The participants inhaled the solution for 9 min while simultaneously preforming breathing exercises in accordance with instructions from the lung clinic at Linköping University Hospital. The participants were asked to cough deeply to expectorate sputum from the lungs. The expectorates were collected into sterile 50 ml Falcon tubes (Thermo Fisher Scientific, Waltham, US) after each session. The Falcon tubes were kept on ice. The sputum inductions were performed in three replicates.
4
Recombinant Protein Production in Pichia pastoris
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Standard laboratory reagents were obtained from Sigma-Aldrich (Vienna, Austria) or Carl Roth GmbH & Co. KG (Karlsruhe, Germany) with the highest purity available. KV and HO-KV were obtained from InnoSyn B.V. (Geleen, The Netherlands). 8-Prenylnaringenin was purchased from Sigma-Aldrich (Vienna, Austria), and Isoxanthohumol was a kind gift of Prof. Michael Murkovic, Institute of Biochemistry, Graz University of Technology. Hop extract capsules were purchased from Allcura (Wertheim, Germany). Restriction enzymes were acquired from Thermo Scientific (St. Leon-Rot, Germany). Bacto™ peptone, Bacto™ yeast extract and Difco™ yeast nitrogen base w/o amino acids (YNB) were obtained from Becton, Dickinson and Company (Schwechat, Austria). ZeocinTM was purchased from InvivoGen (Vienna, Austria). Sterile water was acquired from Fresenius Kabi, Graz, Austria.
P. pastoris cultures were routinely grown in buffered glycerol-complex medium BMGY (1% yeast extract, 2% peptone, 100 mM potassium phosphate, pH 6.0, 1.34% YNB, 4 × 10−5% biotin, 1% glycerol). Buffered methanol-complex medium, BMMY (1% yeast extract, 2% peptone, 100 mM potassium phosphate, pH 6.0, 1.34% YNB, 4 × 10−5% biotin, 1% methanol) was used as induction medium.
P. pastoris cultures were routinely grown in buffered glycerol-complex medium BMGY (1% yeast extract, 2% peptone, 100 mM potassium phosphate, pH 6.0, 1.34% YNB, 4 × 10−5% biotin, 1% glycerol). Buffered methanol-complex medium, BMMY (1% yeast extract, 2% peptone, 100 mM potassium phosphate, pH 6.0, 1.34% YNB, 4 × 10−5% biotin, 1% methanol) was used as induction medium.
5
Sputum Induction with Hypertonic Saline
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The sputum induction 23 (link) was performed with an eFlow rapid nebulizer (PARI, Hamburg, Germany) filled with a hypertonic saline solution. The solution was prepared by mixing sterile water (Fresenius Kabi, Stockholm, Sweden) with 4% sodium chloride (B. Braun Medical AB, Stockholm, Sweden). The participants inhaled the solution for 9 minutes while simultaneously preforming breathing exercises in accordance with instructions from the lung clinic at Linköping University Hospital. The participants were asked to cough deeply to expectorate sputum from the lungs. The expectorates were collected into sterile 50 ml Falcon tubes (Thermo Fisher Scientific, Waltham, US) after each session. The Falcon tubes were kept on ice. The sputum inductions were performed in three replicates.
6
Detailed Cell Culture Protocol
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Minimum Essential Medium (MEM), Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12 (DMEM/F-12), foetal calf serum (FCS) and phosphate buffered saline (PBS) were purchased from Gibco (Cergy-Pontoise, France), non-essential amino acids from Eurobio (Courtaboeuf, France). Trypsin, Giemsa reagent, penicillin, streptomycin, amphotericin B, mitomycin C (MitoC), methyl methanesulfonate (MMS), glucose oxidase (GOx), Triton X-100, EDTA, trizma base, propidium iodide, KCl, NaCl, sodium bicarbonate, sodium pyruvate, and bovine serum albumin (BSA) were obtained from Sigma-Aldrich (Saint-Quentin Fallavier, France). Normal melting point agarose (NMPA) and low melting point agarose (LMPA) were purchased from Biorad (Marnes-la-Coquette, France), acetic acid from VWR (Fontenaysous-Bois, France), human 8-oxoguanine glycosylase (hOGG1) from New England Biolabs (Evry, France), dimethyl sulfoxide (DMSO) from Acros Organics (Noisy-le-Grand, France), NaOH, L-glutamine, methanol and absolute ethanol from Merck (Darmstadt, Germany), and sterile water from Fresenius Kabi (Sèvres, France).
7
Quantification of Drugs in Biological Samples
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Cotinine (COT), trans-3-hydroxycotinine (OH-COT), cotinine-d3 and trans-3-hydroxycotinine-d3 were obtained from LGC Standards (Molsheim, France). Caffeine, ibuprofen, methadone, morphine, pseudoephedrine, codeine, cocaine, clomipramine, propranolol and Δ-9-tetrahydrocannabinol analytical reference standards presented in Methanol were obtained from Cerilliant (Sigma-Aldrich, Saint-Quentin-Fallavier, France). Potassium dihydrogen phosphate and dipotassium hydrogen phosphate were obtained from Sigma-Aldrich (St Louis, MO, USA). Methanol, hexane, isopropanol were of analytical grade and were obtained from Merck (Darmstadt, Germany). Sterile water was obtained from Fresenius (Sevres, France). Ammonia solution 25% was purchased from VWR (Fontenay-sous-Bois, France). Hydrochloric acid 0.2 m was obtained from VWR (Fontenay-sous-Bois, France). Clean Screen SPE columns 200 mg were purchased from UCT (Bristol, PA, USA). The derivatizing agent utilized was bis(trimethylsilyl)trifluoroacetamide (BSTFA) + 1% trimethylchlorosilane (TMS) (Sigma-Aldrich, Saint-Quentin-Fallavier, France).
8
Bioaerosol Sampling and Characterization in Healthcare and Contaminated Homes
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Bioaerosols were collected indoors from a cancer treatment center (Centre François Baclesse, Caen, France) (n = 24) [32 ] and in contaminated homes (n = 65) [18 (link),33 (link)], and outdoor in agricultural environments (silage and hay) (n = 4) [34 (link),35 (link)]. During collection, a Grimm particle counter (Model 1.108, Grimm Technologies, Inc., Douglasville, GA, USA) was used to measure temperature and relative humidity every 6 s. The sampling of bioaerosols in contaminated homes was carried out during two different campaigns: the first one in Serpula lacrymans-damaged homes [33 (link)] and the second in mold-damaged homes [18 (link)].
Samples were cultured on Malt Extract Agar medium with 0.02% chloramphenicol (Cooper, Melun, France) (MEA+). Plates were incubated at 25 °C and checked daily. Each fungal colony was isolated and purified on the same medium. All Aspergillus isolates belonging to the Versicolores series (n = 93) were stored on slant agar at −4 °C and in a cryoprotective agent composed of sterile water (Fresenius Kabi AG, Bad Homburg, Germany) and 10% glycerol (Carlo Erba, Val-de-Reuil, France) at −80 °C before characterization.
Samples were cultured on Malt Extract Agar medium with 0.02% chloramphenicol (Cooper, Melun, France) (MEA+). Plates were incubated at 25 °C and checked daily. Each fungal colony was isolated and purified on the same medium. All Aspergillus isolates belonging to the Versicolores series (n = 93) were stored on slant agar at −4 °C and in a cryoprotective agent composed of sterile water (Fresenius Kabi AG, Bad Homburg, Germany) and 10% glycerol (Carlo Erba, Val-de-Reuil, France) at −80 °C before characterization.
9
Aspergillus Isolation and Characterization
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Bioaerosols were collected in a hospital (Centre François Baclesse, Caen, France) (n = 24) [38] , in Serpula lacrymans-and mold-damaged homes (n = 65) [13] , [39] and outdoor in agricultural environments (silage and hay) (n = 4) [40] , [41] .
Samples were cultured on malt extract agar (MEA) medium with 0.02% chloramphenicol (Cooper, Melun, France). Plates were incubated at 25°C and checked daily. Each fungal colony was isolated and purified on MEA. All Aspergillus isolates belonging to the series Versicolores (n = 93) were stored on slant agar at 4°C and in a cryoprotective agent composed of sterile water (Fresenius Kabi AG, Bad Homburg, Germany) and 10% glycerol (Carlo Erba, Val-de-Reuil, France) at -80°C before molecular characterization.
Samples were cultured on malt extract agar (MEA) medium with 0.02% chloramphenicol (Cooper, Melun, France). Plates were incubated at 25°C and checked daily. Each fungal colony was isolated and purified on MEA. All Aspergillus isolates belonging to the series Versicolores (n = 93) were stored on slant agar at 4°C and in a cryoprotective agent composed of sterile water (Fresenius Kabi AG, Bad Homburg, Germany) and 10% glycerol (Carlo Erba, Val-de-Reuil, France) at -80°C before molecular characterization.
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