2 4 6 trinitrobenzene sulfonic acid tnbsa

Manufactured by Thermo Fisher Scientific

Sourced in Spain, United States, Germany

2,4,6-Trinitrobenzene sulfonic acid (TNBSA) is a chemical reagent commonly used in biochemical assays. It is a yellow crystalline solid that is soluble in water and organic solvents. TNBSA is primarily used as a colorimetric agent for the quantitative determination of primary amino groups in proteins, peptides, and other biomolecules.

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6 protocols using 2 4 6 trinitrobenzene sulfonic acid tnbsa

Functionalized Polymer Hydrogel Synthesis

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Methacrylic acid (MAA), ethylene glycol dimethacrylate (EGDMA), trimethylolpropane trimethacrylate (TRIM), pentaerythritol-tetrakis-(3-mercaptopropionate) (PETMP), phosphate buffer solution (PBS) (pH 7.4), sodium hydroxide, glutaraldehyde (GA), histamine, histidine, melamine, (3-aminopropyl)triethoxysilane (APTES). Carbodiimide (EDC), N-hydroxysuccinimide (NHS) 98%, ethanolamine hydrochloride, hydrochloride acid (HCl), 11-mercaptoundecanoic acid (MUDA) 95%, hydrogen peroxide, acetonitrile (ACN), ethanol, acetone, phthaldialdehyde, 60 mL SPE tubes and 20 μm pore frits and glass beads (75 μm), were purchased from Sigma-Aldrich (DK). 2,4,6-Trinitrobenzene sulfonic acid (TNBSA) was purchased from Thermofisher Scientific (DK). N,N-Diethyldithiocarbamic acid benzyl ester (DABE) was obtained from TCI Europe (Belgium). Q-Max® Celluloseacetat (CA) syringe filters 0.45 μm were from FRISENETTE (DK). Gold chip was purchased from Bionavis, Sweden. Nunclon 96-well flat-bottom black microwell plates were purchased from Thermo scientific, DK. 0.1% milk (Arla) was bought from a local supermarket, Netto. Deionized water was obtained from a Millipore (MilliQ) purification system. All chemicals were analytical or HPLC grade and were used without further purification.

Feng X., Ashley J., Zhou T., Halder A, & Sun Y. (2018). A facile molecularly imprinted polymer-based fluorometric assay for detection of histamine. RSC Advances, 8(5), 2365-2372.

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Hyaluronic Acid Characterization and Antimicrobial Evaluation

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Pharmaceutical grade hyaluronic acid (HA) sodium salt from Streptococcus Equi (MW = 40, 200, 600 kDa and 2 MDa) was obtained from Lehvoss Iberia (Barcelona, Spain). Silver nitrate, Muller-Hinton Broth (MHB), Coliform ChromoSelect agar, Braid-Parker agar, Dulbecco’s modified Eagle’s medium—high glucose (DMEM) were supplied by Sigma-Aldrich (Madrid, Spain). Invitrogen Life Technologies Corporation (Sant Cugat del Vallès, Spain) provided the alamarBlue cell viability reagent. Adipic acid dihydrazide (ADH), hydrochloric acid, nitric acid, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and 2,4,6-trinitrobenzene sulfonic acid (TNBSA) were supplied by Thermo Fischer Scientific (Sant Cugat del Vallès, Spain). Sodium dodecyl sulfate was purchased from Fisher Bioreagent (Madrid, Spain). Bacterial strains Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922), and human fibroblast (ATCC-CRL-4001, BJ-5ta) and keratinocyte (HaCaT cell line) cells were obtained from the American Type Culture Collection (ATCC LGC Standards, Barcelona, Spain).

Ferreres G., Pérez-Rafael S., Torrent-Burgués J, & Tzanov T. (2021). Hyaluronic Acid Derivative Molecular Weight-Dependent Synthesis and Antimicrobial Effect of Hybrid Silver Nanoparticles. International Journal of Molecular Sciences, 22(24), 13428.

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Formulation and Characterization of Nicotine-Conjugated Nanoparticles

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Lactel^® 50:50 PLGA (acid-terminated) was purchased from Durect Corporation (Cupertino, CA, USA). 2,4,6-trinitrobenzenesulfonic acid (TNBSA), Alexa Fluor 350 (AF350), Alexa Fluor 647 (AF647), and keyhole limpet hemocyanin (KLH) were purchased from Thermo Fisher Scientific Inc. (Rockford, IL, USA). 1,2-Dioleoyl-3-trimethylammonium-propane (DOTAP), cholesterol (CHOL), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethylene glycol)-2000] (ammonium salt) (DSPE-PEG2000-maleimide), and 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (ammonium salt) (NBD-PE) were purchased from Avanti Polar Lipids Inc. (Alabaster, AL, USA). O-succinyl-3’-hydroxymethyl-(±)-nicotine (Nic) hapten was purchased from Toronto Research Chemicals (North York, ON, Canada). All other chemicals were of analytical grade.

Zhao Z., Powers K., Hu Y., Raleigh M., Pentel P, & Zhang C. (2017). Engineering of a hybrid nanoparticle-based nicotine nanovaccine as a next-generation immunotherapeutic strategy against nicotine addiction: a focus on hapten density. Biomaterials, 123, 107-117.

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Boc-NH-PEG5000-OH Synthesis and Characterization

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All materials were obtained
from Sigma-Aldrich (Zwijndrecht, The Netherlands) and used as received
unless otherwise noted. α-tert-Butyloxycarbonylamino-ω-hydroxypoly(ethylene
glycol) with PEG molecular weight of 5000 Da (Boc-NH-PEG₅₀₀₀-OH) was obtained from Iris Biotech GmbH (Marktredwitz, Germany)
and dried overnight in a vacuum oven at room temperature (RT) before
use. 4-(Dimethylamino)pyridinium-4-toluenesulfonate (DPTS) was
prepared according to a literature procedure.^{24 (link)} Triethylamine (TEA) and 2,4,6-trinitrobenzenesulfonic acid
(TNBSA) solution in methanol (5% w/v) were obtained from Thermo Fisher
Scientific (Bleiswijk, The Netherlands). Peptide grade dichloromethane
(DCM), N,N-dimethylformamide (DMF), tetrahydrofuran (THF), and diethyl
ether were purchased from Biosolve (Valkenswaard, The Netherlands).
All solvents were dried by molecular sieves for 24 h before use. Prior
to use, 2-(dimethylamino)ethyl methacrylate (DMAEMA) was passed through
a column of alumina to remove the inhibitor. Slide-A-lyzer dialysis
cassettes (M_w cutoff: 3.5 and 10 kDa)
were obtained from Thermo Fisher Scientific (Bleiswijk, The Netherlands).

Fliervoet L.A., Najafi M., Hembury M, & Vermonden T. (2017). Heterofunctional Poly(ethylene glycol) (PEG) Macroinitiator Enabling Controlled Synthesis of ABC Triblock Copolymers. Macromolecules, 50(21), 8390-8397.

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Synthesis and Characterization of Photosensitive Biomaterials

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Aldrithiol-2 and Silica gel (Spherical, 100 μm) were purchased from Tokyo Chemical Industry Co., LTD (Harborgate Street, Portland, OR). 2-mercaptoethenol, DL-dithiothreitol (DTT), tris(2-carboxyethyl)phosphine (TCEP), 2, 2-Azobisisobutyronitrile (AIBN) and Poly(ethylene glycol)methacrylate (Mn = 360 Da) were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO). Penicillin (10,000 U/mL), streptomycin (10,000 μg/mL), 0.25% trypsin-EDTA, Dulbecco’s Modified Eagle Medium (with L-glutamine) and fetal bovine serum (FBS) were obtained from American Type Culture Collection (ATCC, Manassas, VA). 2,4,6-Trinitrobenzene sulfonic acid (TNBSA) was purchased from Thermo Scientific. Silicon phthalocyanine (Pc 4) was acquired from NCI (NSC 676418). All the other solvents used in this research were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO) and used without further purification unless otherwise noted.

He H., Cattran A.W., Nguyen T., Nieminen A.L, & Xu P. (2014). Triple-responsive Expansile Nanogel for Tumor and Mitochondria Targeted Photosensitizer Delivery. Biomaterials, 35(35), 9546-9553.

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Detailed Chemical Inventory for Analytical Assays

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The following chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA): sodium carbonate (≥99%), copper(II) sulfate pentahydrate (≥98%), potassium sodium tartrate tetrahydrate (≥99%), sodium hydroxide (≥97%), Folin-Ciocalteu's phenol reagent, Bovine Serum Albumin (BSA), sodium dodecyl sulfate (SDS), β-mercaptoethanol, bromophenol blue, Coomassie Brilliant Blue (R-250), tetramethylethylenediamine (TEMED), gallic acid (3,4,5 trihydroxybenzoic acid), 2,2 diphenyl-1picrylhydrazyl (DPPH), 2,2 ′ -Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), pepsin from porcine gastric mucosa, αamylase and pancreatin from porcine pancreas, bile extract porcine and pefabloc® SC. Acrylamide/Bis solution (29:1) was purchased from Bio-Rad (California, USA), whereas calcium chloride was obtained from Panreac (Barcelona, Spain). Hydrochloric acid, n-hexane (≥95%), methanol, glacial acetic acid, and 2,4,6 trinitrobenzene sulfonic acid (TNBSA) were purchased from Thermo Fisher Scientific (Bremen, Germany) whereas serine was obtained from Acros Organics (Belgium).

González A., Nobre C., Simões L.S., Cruz M., Loredo A., Rodríguez-Jasso R.M., Contreras J., Texeira J, & Belmares R. (2021). Evaluation of functional and nutritional potential of a protein concentrate from Pleurotus ostreatus mushroom. Food chemistry, 346.

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