Sucrose, fructose and glucose were purchased from Sigma-Aldrich Co. (St. Louis, MO), with purity >99%. Acetic acid (>99% purity), sulfuric acid (>98% purity) and hydrochloric acid (36.5 to 38%) were purchased from Merck Inc. (Emanuel Merck, Darmstadt, Germany). Citric acid (>99% purity) and tartaric acid (>99% purity) were purchased from Mallinckrodt Pharmaceuticals (St. Louis MO) and Spectrum Chemical MFG Corp. (New Brunswick, NJ). Quinine hydrochloride (#Q1125) and denatonium benzoate (#D5765) were purchased form Sigma-Aldrich and lobeline hydrochloride (LTD #L0096) from Tokyo Chemical Industry Co. (Tokyo, Japan). All bitter chemicals were of > 98 % purity.
Citric acid
Manufactured by Mallinckrodt
Citric acid is a chemical compound used in various laboratory applications. It is a weak organic acid found naturally in citrus fruits. Citric acid has a sour taste and is commonly used as a pH regulator, preservative, and flavoring agent in many industries.
Automatically generated - may contain errors
Lab products found in correlation
Sulfuric acid, by Merck Group (2 mentions)
H2so4, by Avantor (2 mentions)
Denatonium benzoate, by Merck Group (1 mentions)
Quinine hydrochloride, by Merck Group (1 mentions)
Glucose, by Merck Group (1 mentions)
Sucrose, by Merck Group (1 mentions)
Fructose, by Merck Group (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Acetic acid, by Merck Group (1 mentions)
Tartaric acid, by Mallinckrodt (1 mentions)
Thiamine, by Thermo Fisher Scientific (1 mentions)
Glucose, by Avantor (1 mentions)
Biotin, by Merck Group (1 mentions)
Mgso4 7h2o, by Thermo Fisher Scientific (1 mentions)
K2hpo4, by Thermo Fisher Scientific (1 mentions)
Kh2po4, by Thermo Fisher Scientific (1 mentions)
Sodium chloride (nacl), by Avantor (1 mentions)
Uv vis spectrometer, by Thermo Fisher Scientific (1 mentions)
Ca oh 2, by Thermo Fisher Scientific (1 mentions)
P aminobenzoic acid, by Thermo Fisher Scientific (1 mentions)
5 protocols using citric acid
1
Bitterness Perception Chemical Preparation
2
Butanol Production from Clostridium acetobutylicum
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Glucose and NaOH were purchased from VWR (West Chester, PA). Ca(OH)2, p-aminobenzoic acid and CH3COONH4 were purchased from Alfa Aesar (Heysham, England). Thiamine was purchased from Alfa Aesar (Ward Hill, MA). Dowex 1X4 resin (chloride form) and biotin were purchased from Sigma-Aldrich (St. Louis, MO). H2SO4 (98%) and NaCl were purchased from VWR (West Radnor, PA). K2HPO4, KH2PO4, MgSO4·7H2O, MnSO4·H2O, FeSO4·7H2O were purchased from Fisher Scientific (Fair Lawn, NJ). Citric acid was purchased from Mallinckrodt Chemicals (Phillipsburg, NJ). CaCO3 was purchased from EMD Chemicals (Gibbstown, NJ). Reinforced Clostridial Broth medium (RCM) was purchased from HIMEDIA laboratories (Mumbai, India). Cellic CTec 2 was obtained from Novozymes North America, Inc (Franklinton, UC). DI water was produced by the Barnstead Nanopure UV Ultrapure Water System (Thermo Fisher Scientific, Marietta, OH).
Clostridium acetobutylicum ATCC 824 was used for butanol production. It was routinely stored as spores at 4 °C and treated by heat shock at 75 °C for 10 min followed by cooling down in an ice bath prior to cultivation. The RCM medium was sparged with nitrogen and then autoclaved at 121 °C for 15 min. The heat-shocked cells were grown until the optical density (OD) reached 1.30 determined by an UV–vis spectrometer (Thermo Scientific, Madison, WI) at 600 nm.
Clostridium acetobutylicum ATCC 824 was used for butanol production. It was routinely stored as spores at 4 °C and treated by heat shock at 75 °C for 10 min followed by cooling down in an ice bath prior to cultivation. The RCM medium was sparged with nitrogen and then autoclaved at 121 °C for 15 min. The heat-shocked cells were grown until the optical density (OD) reached 1.30 determined by an UV–vis spectrometer (Thermo Scientific, Madison, WI) at 600 nm.
3
Comprehensive Intracellular and Extracellular Solutions
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The following chemicals were used in the intracellular and extracellular solutions (for details, see Table 3 and 4 , respectively): CsMeSO3 (C1426), CsCl (289329), CsOH (C8518), NaCl (7548-4400, DAEJUNG), MgCl2 (M9272), MgSO4 (6070-12, Mallinckrodt Chemicals), sodium l -glutamate (49621), KCl (P5405), CaCl2 (C7902), EGTA (E4378), MgATP (A9187), NaGTP (G8877), HEPES (H3375), citric acid (C2404), NaOH (7708-10, Mallinckrodt Chemicals), H2SO4 (339741), HCl (H1758), and d -glucose (1916-01, J.T. Baker). All chemicals were purchased from MilliporeSigma unless otherwise indicated.
4
ELISA for Mouse Antibody Isotypes
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Based on the antigen used for immunizations, NP conjugated to BSA at a >20 ratio (Biosearch Technologies), PE or OVA all diluted in PBS were coated onto separate 96 well plates (Greiner Bio-one) at 2 μg/mL and incubated overnight at 4 °C. Plates were washed then blocked with casein (Thermo Scientific) for one hour at room temperature (RT). Serum samples were serially diluted in casein then added to the plates and incubated for two hours at RT. After washing, biotin rat anti-mouse IgG1 (BD), peroxidase conjugated goat anti-mouse IgM (Jackson Immunoresearch) or biotin rat anti-mouse IgE (BD) was added to the plates and incubated for one hour at RT. Plates were washed and streptavidin HRP (Jackson Immunoresearch) was added to the IgG1 and IgE plates and incubated for thirty minutes at RT. Plates were washed then developed for twenty minutes with an o-Phenylenediamine dihydrochloride tablet (Sigma) dissolved in 100 mM Sodium phosphate dibasic (Mallinckrodt), 50 mM citric acid (Mallinckrodt) and 0.04% hydrogen peroxide (Mallinckrodt) at pH 5.0. Reactions were stopped using 1N HCL (Fisher Scientific) and plates were read at 490λ using a Molecular Devices Spectra Max 340 PC running Soft Max Pro 3.1.2. All antibody data are presented at IgM (1:1000), IgG1 (1:80,000), and IgE (1:100) serum dilutions which represent non-saturating concentrations.
5
Thiolated Oligonucleotides for Graphite-based Biosensors
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All thiolated oligonucleotides used in this study were purchased from Integrated DNA Technologies (Coralville, IA). Graphite powder (99%, 7–11 μM) was purchased from Alfa–Aesar (Heysham, Lancashire, UK). Sulfuric acid, phosphoric acid, hydrogen peroxide, trisodium citrate, sodium phosphate tribasic, sodium phosphate dibasic, boric acid, tetrachloroauric acid, dimethyl sulfoxide, apigenin, xylene, ethanol, 3,3-diaminobenzidine tetrahydrochloride, hematoxylin, and isopropanol were purchased from Sigma–Aldrich (Milwaukee, WI). Citric acid, calcium chloride, magnesium chloride, potassium permanganate, tris(hydroxymethyl)aminomethane (Tris), and hydrochloric acid were purchased from Mallinckrodt Baker (Phillipsburg, NJ). Alpha-MEM and fetal bovine serum were purchased from GIBCO (Campinas, Brazil). An MTT assay kit was purchased from Thermo Fisher Scientific (Waltham, MA). MUC1 (VU4H5) mouse mAb, anti-mouse IgG, and HRP-linked antibody were purchased from Cell Signaling Technology (Danvers, MA). Milli-Q ultrapure water (18 MΩ, Millipore, Billerica, MA) was used in all experiments.
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