EXPAR templates, DNA targets and DNA probe functionalized on AuNPs were purchased from Integrated DNA Technologies. The sequences of DNA used are shown in Table 2 . The Vent (exo-) polymerase, Nt.BtsNBI nicking enzyme, the ThermoPol buffer, the NEBuffer 3.1, BSA, SSB proteins, dNTPs and the Streptavidin magnetic beads, were purchased from New England BioLabs. The Biotin-11-dUTP was obtained from Biotium. Ethylene glycol, propylene glycol, betaine, DMSO, trehalose, TMAC, and HAuCl4, sodium citrate dehydrate, 4-mercaptobenzoic acid (MBA) were purchased from Sigma-Aldrich.
4 mercaptobenzoic acid mba
Manufactured by Merck Group
Sourced in United States
4-mercaptobenzoic acid (MBA) is a common organic compound used in various laboratory applications. It is a white crystalline solid with the chemical formula C6H6O2S. MBA serves as a versatile reagent and building block for chemical synthesis and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Streptavidin magnetic beads, by New England Biolabs (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Bovine serum albumin (bsa), by New England Biolabs (1 mentions)
Propylene glycol, by Merck Group (1 mentions)
Nebuffer 3, by New England Biolabs (1 mentions)
Ethylene glycol, by Merck Group (1 mentions)
Trehalose, by Merck Group (1 mentions)
Haucl4, by Merck Group (1 mentions)
Betaine, by Merck Group (1 mentions)
Thermopol buffer, by New England Biolabs (1 mentions)
Sodium citrate dehydrate, by Merck Group (1 mentions)
Dntps, by New England Biolabs (1 mentions)
Polycarbonate filter, by Cytiva (1 mentions)
1 2 dioleoyl 3 trimethylammonium propane dotap, by Avanti Polar Lipids (1 mentions)
1 palmitoyl 2 oleoyl sn glycero 3 phospho l serine, by Avanti Polar Lipids (1 mentions)
1 palmitoyl 2 oleoyl sn glycero 3 phosphocholine, by Avanti Polar Lipids (1 mentions)
Whatman, by Cytiva (1 mentions)
1 ethyl 3 3 dimethylaminopropyl carbodiimide edc, by Merck Group (1 mentions)
N hydroxysuccinimide nhs, by Merck Group (1 mentions)
Ferrocenecarboxaldehyde, by Merck Group (1 mentions)
7 protocols using 4 mercaptobenzoic acid mba
1
Nanoparticle-Based EXPAR Assay
2
Lipid-Coated Gold Substrates
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Different thiols were used in combination with the different lipids to be coated onto them by electrostatically-driven physisorption. We used two thiols, namely 4-mercaptobenzoic acid (MbA) and 11-amino-1-undecanethiol hydrochloride (AT), from Sigma (Milan, Italy), and three lipids, namely 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (POPS) and, 1,2-dioleoyl-3-trimethylammoniumpropane (DOTAP), from Avanti Polar Lipids (Alabaster, Alabama, US). All solvents were purchased from Sigma-Aldrich.
First, the substrates were incubated at rt for 2 h with a 1 mM aqueous solution of the thiol molecule, either MbA or AT, to let the sulfur of the –SH group bind covalently to the Au surface (chemisorption). The substrates were then gently washed with their aqueous solutions and dried under nitrogen flow.
All the lipids were dissolved in chloroform/methanol 2:1 vol/vol, dried under a gentle nitrogen flux in a test tube, and put under a mild vacuum overnight to remove all solvent traces. POPC/POPS in a 9:1 mol/mol ratio and DOTAP were then re-suspended in PBS at a 5 g/L concentration, let to swell for 30 min, and extruded 11 times through a polycarbonate filter (Whatman, USA) with 100 nm pore diameter to form unilamellar vesicles.
First, the substrates were incubated at rt for 2 h with a 1 mM aqueous solution of the thiol molecule, either MbA or AT, to let the sulfur of the –SH group bind covalently to the Au surface (chemisorption). The substrates were then gently washed with their aqueous solutions and dried under nitrogen flow.
All the lipids were dissolved in chloroform/methanol 2:1 vol/vol, dried under a gentle nitrogen flux in a test tube, and put under a mild vacuum overnight to remove all solvent traces. POPC/POPS in a 9:1 mol/mol ratio and DOTAP were then re-suspended in PBS at a 5 g/L concentration, let to swell for 30 min, and extruded 11 times through a polycarbonate filter (Whatman, USA) with 100 nm pore diameter to form unilamellar vesicles.
3
Enzyme-based Biosensing Platform Development
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Organic solvents were purchased from ChemPur (Piekary Śląskie, Poland) and distilled prior to the experiments. The used lyophilized powder enzymes of type VII glucose oxidase (GOx) from Aspergillus niger of ≥100 U mg−1 (without added oxygen) and type I peroxidase from horseradish (HRP) of approximately 150 U mg−1 were purchased from Sigma Aldrich. All inorganic salts for the buffer solutions were purchased from Carl Roth GmbH + Co. KG. Ferrocenecarboxaldehyde, 4-mercaptobenzoic acid (MBA), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS) were purchased from Sigma Aldrich. All the experiments were conducted using type II water (R > 18 MΩ) purified in a Milli-Q system; 50 mM pH 7.0 potassium phosphate buffer (PPB) solution was used.
4
Fabrication of SERS Microspheres for Sensing
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SERS-MS were
fabricated using 20 μm TentaGel M NH2 microspheres
(Rapp Polymere GmbH, M30202, 2.4 × 108 particles/g)
as previously described.9 (link) TentaGel M NH2 powder (1 mg) was added to 3 mL of citrate-capped 150 nm
gold nanoparticle colloid (Sigma-Aldrich, 746649), sonicated for 10
min, and stored at 4 °C. The day before sensing experiments,
the particles were pelleted via centrifugation and suspended in a
100 μM solution of 4-mercaptobenzoic acid (MBA) (Sigma-Aldrich,
662534) in 1% EtOH (≥99.8%, Sigma-Aldrich) in deionized (DI)
water. The MBA solution was prepared by dissolving MBA in EtOH at
10 mM and diluting 1:100 with DI water. The SERS-MS were stored in
the MBA solution overnight at 4 °C before washing with 70% EtOH
in DI water and twice with sterile saline solution. The SERS-MS were
suspended at a final concentration of 2000 SERS-MS/μL in sterile
saline before application to ALI cultures. For scanning electron microscope
(SEM) imaging, SERS-MS was suspended in EtOH and dried onto an aluminum
stub. SEM images were collected with a Zeiss Crossbeam 550.
fabricated using 20 μm TentaGel M NH2 microspheres
(Rapp Polymere GmbH, M30202, 2.4 × 108 particles/g)
as previously described.9 (link) TentaGel M NH2 powder (1 mg) was added to 3 mL of citrate-capped 150 nm
gold nanoparticle colloid (Sigma-Aldrich, 746649), sonicated for 10
min, and stored at 4 °C. The day before sensing experiments,
the particles were pelleted via centrifugation and suspended in a
100 μM solution of 4-mercaptobenzoic acid (MBA) (Sigma-Aldrich,
662534) in 1% EtOH (≥99.8%, Sigma-Aldrich) in deionized (DI)
water. The MBA solution was prepared by dissolving MBA in EtOH at
10 mM and diluting 1:100 with DI water. The SERS-MS were stored in
the MBA solution overnight at 4 °C before washing with 70% EtOH
in DI water and twice with sterile saline solution. The SERS-MS were
suspended at a final concentration of 2000 SERS-MS/μL in sterile
saline before application to ALI cultures. For scanning electron microscope
(SEM) imaging, SERS-MS was suspended in EtOH and dried onto an aluminum
stub. SEM images were collected with a Zeiss Crossbeam 550.
5
Synthesis and Characterization of Europium-doped Nanoparticles
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Calcium nitrate tetrahydrate, (Ca(NO3)2.4H2O, 99%, Alfa Aesar), ammonium molybdate (H8MoN2O4, 99.99%, Alfa Aesar, Ward Hill, MA, USA), europium(III) nitrate hydrate (Eu(NO3)3 .xH2O, 99.99%, Sigma Aldrich, St. Louis, MO, USA), oleic acid (Alfa Aesar), 1-octadecene (95%, Alfa Aesar), NaOH pellet (Merck, Kenilworth, NJ, USA), hydrochloric acid (HCl, Sigma-Aldrich), HS-PEG-COOH (MW = 5000, NANOCS, New York, NY, USA), mPEG-SH (MW = 5000, NANOCS), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (C8H17N3HCl, MW = 191.7 g mol–1, Sigma-Aldrich) (EDC), N-hydroxysuccinimide (C4H5NO3, Sigma-Aldrich, MW = 115.09 g mol–1) (NHS), 4-mercaptobenzoic acid (MBA) (Sigma-Aldrich), anti-human epidermal growth factor receptor (EGFR) antibody (Thermo Fisher Scientific, Waltham, MA, USA), and phosphate buffered saline (1×) (PBS) (Thermo Fisher Scientific) were used for HNP synthesis. Human (Homo sapiens) lung carcinoma (A549 cells) (ATCC, USA), mouse hepatocyte cells (AML12, normal hepatocyte from liver tissue), ethylenediaminetetraacetic acid (EDTA) stabilized human whole blood was freshly obtained from Innovative Research (Novi, MI, USA), 0.5% trypsin-EDTA solution (Life Technologies), LIVE/DEAD viability/cytotoxicity assay kit (Life Technologies), Earle’s balanced salt solution (EBSS) (Life Technologies) and PBS were used for cell experiments.
6
Synthesis and Characterization of Gold Nanoparticles
7
Influenza Virus Binding Assay
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Sodium hydroxide (≥98% pure), HEPES
(≥99.5% pure), and 4-mercaptobenzoic acid (MBA) were purchased
from Sigma-Aldrich. Water used for the preparation of all solutions
was purified with the PURELAB Chorus purification system (Veolia Water
Technologies, Saint-Maurice. France). Recombinant N2 NA with Histidine
Tag from Influenza A/canine/Illinois/11613/2015 (H3N2) (FR-1479),
BPL-Inactivated Influenza A Virus, A/Anhui/01/2005(H5N1)-PR8-IBCDC-RG6
(FR-918), BPL-Inactivated Influenza A, A/Michigan/45/2015 (H1N1)pdm09
(FR-1506), BPL-Inactivated Influenza A Virus, A/Alaska/232/2015 (H3N2)
(FR-1541), and BPL-Treated Influenza A, A/Anhui/1/2013 (H7N9) (FR-1283)
were obtained from the International Reagent Resource, Influenza Division,
WHO Collaborating Center for Surveillance, Epidemiology and Control
of Influenza, Centers for Disease Control and Prevention, Atlanta,
GA, USA. E2-OH, E4-SA, and E4-Zan were synthesized as described in
our previous reports19 (link),21 (link) and in theSupporting Information (E4-Zan).
(≥99.5% pure), and 4-mercaptobenzoic acid (MBA) were purchased
from Sigma-Aldrich. Water used for the preparation of all solutions
was purified with the PURELAB Chorus purification system (Veolia Water
Technologies, Saint-Maurice. France). Recombinant N2 NA with Histidine
Tag from Influenza A/canine/Illinois/11613/2015 (H3N2) (FR-1479),
BPL-Inactivated Influenza A Virus, A/Anhui/01/2005(H5N1)-PR8-IBCDC-RG6
(FR-918), BPL-Inactivated Influenza A, A/Michigan/45/2015 (H1N1)pdm09
(FR-1506), BPL-Inactivated Influenza A Virus, A/Alaska/232/2015 (H3N2)
(FR-1541), and BPL-Treated Influenza A, A/Anhui/1/2013 (H7N9) (FR-1283)
were obtained from the International Reagent Resource, Influenza Division,
WHO Collaborating Center for Surveillance, Epidemiology and Control
of Influenza, Centers for Disease Control and Prevention, Atlanta,
GA, USA. E2-OH, E4-SA, and E4-Zan were synthesized as described in
our previous reports19 (link),21 (link) and in the
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