Soluble biotinylated CD1B monomers were provided by the National Institutes of Health Tetramer Core Facility (Emory University, Atlanta, GA). Glucose monomycolate (GMM)-loaded tetramers were generated as previously described (18 (link)). In brief, C32-GMM was dried down in a glass tube using a nitrogen evaporator and sonicated into 0.25% CHAPS/sodium citrate at pH 4 (preparation of CHAPS in sodium citrate; CHAPS Hydrate, Sigma; Sodium Citrate Dihydrate, Fisher) for two minutes at 37°C. The lipid solution was transferred to a microfuge tube, and 9 μL of CD1B monomer was added. The CD1B-GMM preparation was then incubated in a 37°C water bath for 2 hours with vortexing every 30 minutes. At the end of the incubation, the solution was neutralized to pH 7.4 with 6 μL of 1M Tris pH 9. Finally, 10 μL of Streptavidin conjugated to allophycocyanin (APC) or phycoerythrin (PE) (Life Technologies) was added in ten aliquots of 1 μL every 10 minutes to facilitate tetramerization. The final product was filtered through a SpinX column (Sigma) to remove aggregates and stored at 4°C until use.
Sodium citrate dihydrate
Manufactured by Thermo Fisher Scientific
Sourced in United States
Sodium citrate dihydrate is a chemical compound that is commonly used in laboratory settings. It is a white, crystalline solid that is soluble in water. Sodium citrate dihydrate is a salt of citric acid and is primarily used as a buffering agent to maintain a specific pH range in various laboratory applications.
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Lab products found in correlation
Silver nitrate agno3, by Merck Group (2 mentions)
Sodium hydroxide (naoh), by Merck Group (2 mentions)
Hydrochloric acid (hcl), by Thermo Fisher Scientific (2 mentions)
Sodium borohydride, by Merck Group (2 mentions)
Chloroauric acid trihydrate, by Merck Group (2 mentions)
L ascorbic acid, by Merck Group (2 mentions)
2 thiouracil, by Merck Group (2 mentions)
Ammonium hydroxide, by Thermo Fisher Scientific (2 mentions)
Spinx column, by Merck Group (1 mentions)
Chaps hydrate, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Triton x 100, by Avantor (1 mentions)
Tween 20, by Merck Group (1 mentions)
Haucl4, by Merck Group (1 mentions)
Lipoic acid, by Merck Group (1 mentions)
Hepes, by Avantor (1 mentions)
Irgacure 2959, by Merck Group (1 mentions)
Ph test strips, by Merck Group (1 mentions)
Polyethylene glycol sorbitan monolaurate, by Merck Group (1 mentions)
I2959, by Merck Group (1 mentions)
15 protocols using sodium citrate dihydrate
1
Generating CD1B-GMM Tetramers
2
Aqueous Synthesis of Gold and Silver Nanoparticles
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Milli-Q water was freshly prepared using a Barnstead NANOpure II water filtration system. for z. All nanoparticle reactions were performed in an aqueous environment. Irgacure-2959 ([2-Hydroxy-4′-(2-hydroxyethoxy)-2-methylpropiophenone], 98%, Sigma-Aldrich) stock was prepared at a concentration of 10 mM and used as a photoinitiated reducing electron source. HAuCl4 ⋅ xH2O (chloroauric acid, 50% basis, Sigma-Aldrich) was used as an aqueous gold source and AgNO3 (silver nitrate, 99%, Sigma-Aldrich) as a silver source.
Tween-20 (polyethylene glycol sorbitan monolaurate, ~ 20n, Sigma-Aldrich), sodium citrate dihydrate (99.9%, Fisher), Triton-X 100 (~ 10n, VWR)), HEPES (N-[2-Hydroxyethyl] piperazine-N’-[2-ethanesulfonic acid], 99.5%, Sigma), lipoic acid (98%, Sigma), CLKRS peptide (95%), and bovine serum albumin (98%, Sigma) were used as stabilizing nanoparticle capping agents. Gold nanoparticles were synthesized at a concentration 1 mM and then diluted with solution containing capping agent to 0.66 mM while silver nanoparticles were synthesized at 0.66 mM. After mixing the base reagents (water, I-2959, Tween-20, HAuCl4) pH was ~ 5 and after the reaction pH decreased to ~ 2 as measured by pH test strips (Sigma-Aldrich).
Tween-20 (polyethylene glycol sorbitan monolaurate, ~ 20n, Sigma-Aldrich), sodium citrate dihydrate (99.9%, Fisher), Triton-X 100 (~ 10n, VWR)), HEPES (N-[2-Hydroxyethyl] piperazine-N’-[2-ethanesulfonic acid], 99.5%, Sigma), lipoic acid (98%, Sigma), CLKRS peptide (95%), and bovine serum albumin (98%, Sigma) were used as stabilizing nanoparticle capping agents. Gold nanoparticles were synthesized at a concentration 1 mM and then diluted with solution containing capping agent to 0.66 mM while silver nanoparticles were synthesized at 0.66 mM. After mixing the base reagents (water, I-2959, Tween-20, HAuCl4) pH was ~ 5 and after the reaction pH decreased to ~ 2 as measured by pH test strips (Sigma-Aldrich).
3
Synthesis and Characterization of Gold Nanoparticles
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Gold chloride
(99%), sodium citrate dihydrate, tetrahydrofuran (THF), Ce6, ethanol,
and phosphate buffer saline (PBS, 10× solution) were purchased
from Fisher Scientific. Trypticase soy broth (TSB) and Mueller Hinton
II broth were purchased from Becton Dickinson. S. aureus (ATCC BAA-44) and MCF-7 cell line (ATCC HTB-22) were purchased from
the American Type Culture Collection (ATCC, USA). N-Isopropylacrylamide (NIPAAm, 99%), 2,2′-azobis(2-methylpropionitrile)
(99%), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
were purchased from Sigma-Aldrich. Sodium borohydride was purchased
from Fluka. All chemicals were used as received.
(99%), sodium citrate dihydrate, tetrahydrofuran (THF), Ce6, ethanol,
and phosphate buffer saline (PBS, 10× solution) were purchased
from Fisher Scientific. Trypticase soy broth (TSB) and Mueller Hinton
II broth were purchased from Becton Dickinson. S. aureus (ATCC BAA-44) and MCF-7 cell line (ATCC HTB-22) were purchased from
the American Type Culture Collection (ATCC, USA). N-Isopropylacrylamide (NIPAAm, 99%), 2,2′-azobis(2-methylpropionitrile)
(99%), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
were purchased from Sigma-Aldrich. Sodium borohydride was purchased
from Fluka. All chemicals were used as received.
4
Bacterial Culture and Simulated Digestion
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For bacterial culture preparation, MRS broth was obtained from Becton Dickinson and Company (Sparks, MD, USA). Agar, L -cysteine hydrochloric acid, and sodium chloride (NaCl) were purchased from Sigma–Aldrich (St. Louis, MO, USA). Glycerol and sodium citrate dihydrate was purchased from Fisher Scientific (Fair Lawn, NJ, USA).
For encapsulation experiments, sodium alginate (TICA-algin HG 400 powder) was donated by TIC Gums (White Marsh, ML, USA). Calcium chloride hexahydrate, chitosan (medium molecular weight) was obtained from Sigma-Aldrich. Glacial acetic acid was purchased from Fisher Scientific.
For simulated digestion, ammonium nitrate, bile extract porcine, lipase from porcine pancreas type II, pepsin from porcine gastric mucosa, porcine gastric mucin type II, potassium chloride, potassium citrate, potassium phosphate, sodiumDL -lactate, sodium hydroxide (NaOH), and uric acid sodium salt were also purchased from Sigma-Aldrich. Hydrochloric acid (HCl), phosphate buffer saline (PBS), and urea were purchased from Fisher Scientific.
For encapsulation experiments, sodium alginate (TICA-algin HG 400 powder) was donated by TIC Gums (White Marsh, ML, USA). Calcium chloride hexahydrate, chitosan (medium molecular weight) was obtained from Sigma-Aldrich. Glacial acetic acid was purchased from Fisher Scientific.
For simulated digestion, ammonium nitrate, bile extract porcine, lipase from porcine pancreas type II, pepsin from porcine gastric mucosa, porcine gastric mucin type II, potassium chloride, potassium citrate, potassium phosphate, sodium
5
Synthesis of Gold Nanoparticles
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Chloroauric acid trihydrate (HAuCl4·3H2O, 99.9%), L-ascorbic acid (C6H8O6, ≥99%), sodium borohydride (NaBH4, ≥98.0%), and 2-thiouracil (C4H4N2OS, 97%) were purchased from Sigma-Aldrich, St. Louis, MO, USA, and were used without further modifications. Sodium citrate dihydrate (C6H5Na3O7·2H2O, >95%), dodecyl sulfate (C12H25NaO4S, ≥99%), hydrochloric acid (HCl, 37%), and nitric acid (HNO3, 70%) were obtained from Fisher Scientific, Pittsburgh, PA, USA. Ultrapure water (~18 MΩ·cm) was utilized for all syntheses and solution preparations. All glassware and the magnetic stir bars were cleaned with aqua regia solution (HCl: HNO3 volumetric ratio of 3:1) and rinsed with ultrapure water before use.
6
Characterization of Methylobacterium radiotolerans
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Low viscosity sodium alginate (A1112), Schiff's fuchsin sulfite reagent, sodium metabisulfite, periodic acid, glutamic acid, and phosphate buffered saline (PBS) were purchased from Sigma-Aldrich (St. Louis, MO). Dicalcium phosphate dihydrate, sodium citrate dihydrate, succinic acid, ammonium hydroxide, SYTO9 nucleic acid stain, propidium iodide, and isopropanol were purchased from Fisher (Fair Lawn, NJ). Bacto soytone was purchased from BD Difco BBL (Houston, TX). MilliQ water with a minimum resistivity of 18 MΩ-cm (Millipore, Billerica, MA) was used for all experiments.
The pink-pigmented methylobacteria used in this study were provided by NewLeaf Symbiotics Inc. (St Louis, MO). The strain is deposited at the Northern Regional Research Labs as NRRL Accession #B50930. Based on MALDI-TOF and 16S rRNA sequence (Accugenix® Charles River, Wilmington MA), the isolate was designated as Methylobacterium radiotolerans. Whole genome sequence information (Ilumina Hi-Seq, MOgene Dx, St Louis, MO and Pacific Biosciences, Menlo Park), coupled with ANI score analysis suggests that this strain is closely related to the type strain designated as M. radiotolerans.
The pink-pigmented methylobacteria used in this study were provided by NewLeaf Symbiotics Inc. (St Louis, MO). The strain is deposited at the Northern Regional Research Labs as NRRL Accession #B50930. Based on MALDI-TOF and 16S rRNA sequence (Accugenix® Charles River, Wilmington MA), the isolate was designated as Methylobacterium radiotolerans. Whole genome sequence information (Ilumina Hi-Seq, MOgene Dx, St Louis, MO and Pacific Biosciences, Menlo Park), coupled with ANI score analysis suggests that this strain is closely related to the type strain designated as M. radiotolerans.
7
Synthesis and Characterization of Metallic Nanostructures
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Gold(III) chloride trihydrate (HAuCl4·3H2O, 99.9%), palladium(II) chloride solution
(H2PdCl4), chloroplatinic acid solution (H2PtCl6), hexadecyltrimethylammonium bromide (CTAB,
99%), 4-nitrothiolphenol (4-NTP), 4-MBA, 2-nitro-5-sulfanylbenzoic
acid, sodium hydroxide (NaOH, 98%), potassium iodide (KI, 99%),l -ascorbic acid (AA, 99%), and sodium borohydride (NaBH4, 99%) were purchased from Sigma-Aldrich (St. Louis, MO).
Sodium citrate dihydrate (Na-Cit, 99%) was purchased from Fisher scientific
(Waltham, MA). Ethanol was purchased from Decon Laboratories (King
of Prussia, PA). Silver nitrate (AgNO3) and poly(vinylpyrrolidone)
(PVP, MW ≈ 29,000, 0.7 mM in terms
of the repeating unit) were purchased from Aldrich. All chemicals
were used as received without purification.
(H2PdCl4), chloroplatinic acid solution (H2PtCl6), hexadecyltrimethylammonium bromide (CTAB,
99%), 4-nitrothiolphenol (4-NTP), 4-MBA, 2-nitro-5-sulfanylbenzoic
acid, sodium hydroxide (NaOH, 98%), potassium iodide (KI, 99%),
Sodium citrate dihydrate (Na-Cit, 99%) was purchased from Fisher scientific
(Waltham, MA). Ethanol was purchased from Decon Laboratories (King
of Prussia, PA). Silver nitrate (AgNO3) and poly(vinylpyrrolidone)
(PVP, MW ≈ 29,000, 0.7 mM in terms
of the repeating unit) were purchased from Aldrich. All chemicals
were used as received without purification.
8
Synthesis and Characterization of Gold Nanoparticles
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Chloroauric acid trihydrate (HAuCl4·3H2O) >99.9% and 2-thiouracil (C4H4N2OS) 97% were purchased from Sigma-Aldrich and were used without further modifications. Sodium citrate dihydrate (C6H5Na3O7·2H2O) >95% was obtained from Fisher Scientific. Ultrapure water (>17 MΩ) was utilized for all syntheses and solution preparations. All glassware and the magnetic stir bars were cleaned with aqua regia solution (HCl/HNO3 = 3:1) and rinsed with ultrapure water before use. For cell culture studies, the MDA-MB-231 cancer cell line was obtained from the American Type Culture Collection (ATCC HTB-26).
9
Cream Lipid Extraction and Analysis
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No human or animal subjects were used, so this study did not require approval by an Institutional Animal Care and Use Committee or Institutional Review Board. Raw cream (approximately 40% fat) was obtained from a local dairy plant (Quebec, QC, Canada). Sodium citrate dihydrate, ethyl ether, petroleum ether, chloroform, and methanol were all provided by Fisher Chemical. Sodium azide, ammonium hydroxide, HCl, and phenolphthalein were obtained from Thermo Fisher Scientific. Nitric acid was purchased from Anachemia, and 95% ethanol from Commercial Alcohols. Ammonium thiocyanate and iron(III) chloride hexahydrate were supplied by Sigma-Aldrich, 45-µm syringe filters by Chromatographic Specialties Inc., and grade 1 and grade 5 qualitative filter paper (12.5 mm) by Whatman. All of the following were purchased from BioRad: Mini-PROTEAN TGX Stain-Free Gels (12%, 15-well comb, 15 µL), Laemmli sample buffer, Precision Plus Protein All Blue Standards, and 10 × Tris/Glycine/SDS buffer. Fast Green FCF, Nile Red, and 2-mercaptoethanol were supplied by Sigma-Aldrich.
10
Synthesis of Nanostructured Materials
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All chemicals were purchased from Sigma-Aldrich
(St. Louis, MO), with the exception of sodium citrate dihydrate (≥99.0%)
obtained from Thermo Fisher Scientific (Waltham, MA), ammonium hydroxide
(28.0–30.0%) from J.T. Baker (Center Valley, PA), and poly(ethylene
glycol) methyl ether thiol (mPEG thiol, 500 MW), which was purchased
from Rapp Polymere GmbH (Tübingen, Germany). The following
chemicals were ordered from Sigma-Aldrich: hydrogen tetrachloroaurate(III)
trihydrate (99.9%), hydroxylamine hydrochloride (≥96.0%), (3-aminopropyl)triethoxysilane
(APTES, 99.0%), 2-propanol (≥99.5%), ethanol (≥99.5%),
sodium silicate (reagent grade), tetraethyl orthosilicate (TEOS, ≥99.0%),
1-octadecene (90.0%), trioctylphosphine oxide (99.0%), trioctylphosphine
(97.0%), oleylamine (70.0%), 1-octanethiol (>98.5%), selenium dioxide
(SeO2, 99.9%), sulfur powder (99.9%), poly(methyl methacrylate)
(350k MW), cadmium oxide (CdO, 99.9%), cadmium nitrate tetrahydrate
(Cd(NO3)2·4H2O, 99.9%), selenium
powder (Se, 99.9%), myristic acid (99.0%), oleic acid (90.0%) and
octadecylphosphonic acid (97.0%).
(St. Louis, MO), with the exception of sodium citrate dihydrate (≥99.0%)
obtained from Thermo Fisher Scientific (Waltham, MA), ammonium hydroxide
(28.0–30.0%) from J.T. Baker (Center Valley, PA), and poly(ethylene
glycol) methyl ether thiol (mPEG thiol, 500 MW), which was purchased
from Rapp Polymere GmbH (Tübingen, Germany). The following
chemicals were ordered from Sigma-Aldrich: hydrogen tetrachloroaurate(III)
trihydrate (99.9%), hydroxylamine hydrochloride (≥96.0%), (3-aminopropyl)triethoxysilane
(APTES, 99.0%), 2-propanol (≥99.5%), ethanol (≥99.5%),
sodium silicate (reagent grade), tetraethyl orthosilicate (TEOS, ≥99.0%),
1-octadecene (90.0%), trioctylphosphine oxide (99.0%), trioctylphosphine
(97.0%), oleylamine (70.0%), 1-octanethiol (>98.5%), selenium dioxide
(SeO2, 99.9%), sulfur powder (99.9%), poly(methyl methacrylate)
(350k MW), cadmium oxide (CdO, 99.9%), cadmium nitrate tetrahydrate
(Cd(NO3)2·4H2O, 99.9%), selenium
powder (Se, 99.9%), myristic acid (99.0%), oleic acid (90.0%) and
octadecylphosphonic acid (97.0%).
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