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3 3 5 5 tetramethylbenzidine (tmb)

Sourced in China

3,3',5,5'-tetramethylbenzidine is a colorimetric substrate commonly used in enzyme-linked immunosorbent assays (ELISAs) and other bioanalytical applications. It undergoes an oxidation reaction in the presence of certain enzymes, producing a blue colored product that can be measured spectrophotometrically.

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20 protocols using 3 3 5 5 tetramethylbenzidine (tmb)

1

Synthesis of Plasmonic Gold Nanostructures

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All chemicals were commercially available and used without further purification. Custom-made DGEAGC polypeptides were obtained from Bankpeptide Biological Technology Co., Ltd, (Hefei, China). Hydrogen tetrachloroaurate tetrahydrate (HAuCl4·4H2O, ≥99.99%, metals basis) was purchased from Sino-Platinum Metals Co., Ltd. (Shanghai, China). Hydrochloric acid (HCl, ≥98%); and sodium hydroxide (NaOH, ≥97%) were purchased from Sinopharm Group Co., Ltd, (Beijing, China). Hydrogen peroxide solution(H2O2, ≥30%) was purchased from Xilong Science Co., Ltd, (Shanghai, China). The 3,3’,5,5’-tetramethylbenzidine (TMB, ≥99%) was purchased from Shanghai Macklin Biochemical Co., Ltd, (Shanghai, China). Terephthalic acid (TA, ≥98%) was purchased from Shanghai Macklin Biochemical Co., Ltd. All glassware was thoroughly cleaned with aqua regia (HCl/HNO3 3/1 v/v), rinsed with copious amounts of pure water, and then dried in an oven prior to use.
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2

Synthesis of Gold Nanoparticles with Cyclodextrins

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Hydrogen tetrachloroaurate (III) trihydrate (99.9%) was purchased from Sigma-Aldrich (Milwaukee, WI, USA). α-cyclodextrin (α-CD), β-cyclodextrin (β-CD) and γ-cyclodextrin (γ-CD) were acquired from Aladdin (Shanghai, China). Sodium borohydride (NaBH4), 1-adamantane carboxylic acid, 3,3′,5,5′-tetramethylbenzidine (TMB), skim milk, 1-Ethyl-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide sodium (NHS) were purchased from Macklin (Shanghai, China). All chemicals were of analytical grade.
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3

Lateral Flow Assay Fabrication Protocol

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LiF (98%), Ti3AlC2 (98%), H2PtCl6·6H2O (Pt ≥ 37.5%) and 3,3′,5,5′-tetramethylbenzidine (TMB) were purchased from Macklin Inc. (Macklin, Shanghai, China). Bovine serum albumin (BSA, 98%), sodium borohydride (NaBH4) and 3-amino-9-ethylcarbazole (AEC) were bought from Sigma-Aldrich (St. Louis, MO, USA). The polyclonal Ab against CAP (anti-CAP Ab, 50 mg/mL), goat anti-rabbit IgG (17.5 mg/mL) and coating Ag (CAP-BSA, 15 mg/mL) were prepared in our lab. The sample pads (SB08, GF-2), absorbent pads (CH37K) and polyvinyl chloride (PVC) gasket (SMA31-40) were bought from Shanghai Liangxin Co., Ltd. (Shanghai, China). The nitrocellulose membrane (UniSart CN95) was obtained from Sartorius Stedim Biotech GmbH (Goettingen, Germany).
The BioDot-XYZ3060 dispensing platform was made by BioDot Corporation (Irvine, CA, USA). The high-speed refrigerated centrifuge (GL-23M) was purchased from Hunan Xiangyi Technology Co., Ltd. (Changsha, China). The CTS300 automatic slitting machine and programmable ZQ-2000 strip cutting machine were manufactured by Shanghai Jinbiao Biotechnology, Inc. (Shanghai, China).
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4

Ultrasound-Assisted Synthesis of Rare-Earth Nanomaterials

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All ILs were purchased from Kekaite Co. (Lanzhou, China). Reagent grade depleted Pr(NO3)3 and 3,3,5,5-tetramethylbenzidine (TMB) were obtained from Macklin Co. Ltd. (Shanghai, China). The trans-resveratrol was purchased from Solarbio Life Science (Beijing, China). All other chemicals, including hydrazine hydrate, ethylenediamine, H2O2 and ethanol, were of analytical grade and used without further purification.
Ultrasound synthesis was assisted by an ultrasonic homogenizer (220 V, 950 W) (Ningbo Scientz Biotechnology Co., Ltd.). Powder X-ray diffraction (XRD) analyses were performed on a Bruker D8 Advance diffractometer with Cu Kα radiation, and scanning electron microscopy (SEM) observations were performed with a Hitachi S-4800 scanning electron microscope, and the morphology and sizes of as-prepared materials were analyzed with JEOL 2100 high resolution transmission electron microscope (HRTEM) (JEOL, Japan). An EMGA-920 Oxygen Elemental Analyzer (HORIBA, Japan) was used to determine the oxygen content. The chemical composition of the products was examined with X-ray photoelectron spectrometer (XPS, ESCALAB 250Xi, ThermoFisher). UV-Visible absorption spectra were recorded on a UV-A390 spectrometer (AOE Instruments, Shanghai, China) with 1.0 cm path length.
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5

Multifunctional Antimicrobial Hydrogel Synthesis

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CoCl2·6H2O, tannin, silver acetate, 3,3',5,5'-tetramethylbenzidine (TMB), o-phenylenediamine (OPD) and gelatin were obtained from Macklin Biochemical Co., Ltd. (Shanghai, China). Acrylic anhydride was purchased from Sigma (St Louis, MO, USA). Hydrogen peroxide (H2O2, 30%), aqueous ammonia, acetic acid and ethanol were the products of Tianjin Kermel Chemical Plant (Tianjin, China). 2,2-Dimethoxy-2-phenylacetophenone (DMPA), the live/dead (SYTO 9/PI) bacterial viability kit and PE anti-mouse CD45 (Cat# 12-0451-82) were received from Thermo Fisher Scientific (New Jersey, USA). The lactate dehydrogenase (LDH) cytotoxicity assay kit was obtained from Beyotime (Shanghai, China). 2',7'-Dichlorodihydrofluorescein diacetate (DCFH-DA) was obtained from Solarbio Life Science (Beijing, China). FITC anti-mouse F4/80 (Cat# 123108) and FITC anti-mouse Ly-6C/Ly-6C (Gr-1) (Cat# 108406) were obtained from Biolegend (San Diego, CA, USA). Anti-Ly6G + Ly6c (ab25377) and anti F4/80 (ab111101) were purchased from Abcam (Cambridge, UK). GTVisionTM I detection system/Mo&Rb (GK500510A) was received from Gene Tech (Shanghai) company limited. Four strains of MDR P. aeruginosa (21027, 20059, 21173, 21715) were kindly provided by Qingdao Municipal Hospital (China). Doubly deionized water was purified by a Unique-R20 purification system (Xiamen, China).
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6

Versatile Nanodiamond-based Analytical Platform

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Nano-diamond, FeCl3, H2SO4 and HNO3, 3,3′,5,5′-tetramethylbenzidine (TMB), 1,2-diaminobenzene (OPD), 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS), L-γ-glutamyl-L-cysteinylglycine (GSH), thiourea were purchased from Macklin reagent Co.,Ltd. (Shanghai, China), Human serum albumin (HAS), Bovine serum albumin (BSA), ascorbic acid (AA), glycine (Gly), L-lysine (Lys), L-serine (Ser), D/L-cysteine(D-Cys, L-Cys), glucose, sodium chloride, calcium chloride, cupric sulfate, ferric chloride, potassium chloride, zinc sulfate, magnesium sulfate, tartaric acid, choline chloride were purchased from Sinopharm Chemical Reagent Co. Ltd. NaN3 was purchased fromTianjin Fuchen Chemical Reagent Factory. H2O2 was purchased from Xilong Scientific Co., Ltd. sodium chloride, calcium chloride, cupric sulfate, ferric chloride, potassium chloride, zinc sulfate, magnesium sulfate, tartaric acid, choline chloride was purchased fromSinopharm ChemicalReagent Co., Ltd. Sodium acetate buffer (0.1 M, pH = 4.0) were freshly prepared before use. All solutions prepared for purified water are derived from Wahaha purified water (China).
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7

Fabricating Manganese Chloride Nanoparticles

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Anhydrous manganese chloride (MnCl2) and O-phenylenediamine were purchased from Aladdin Reagents Co., Ltd. (Shanghai, China). Sodium hydroxide (NaOH) and zinc acetate (ZnAc) were acquired from the National Pharmaceutical Group Chemical Reagents Co., Ltd. (Shanghai, China). Bovine serum albumin (BSA) Fraction V and 3,3′,5,5′-tetramethylbenzidine were received from Shanghai Macklin Biochemical Co., Ltd. (Shanghai, China). N-(3-(Dimethylamino)-propyl)-N′-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) were purchased from Sigma-Aldrich (USA). All other chemicals and reagents used were of analytical grade and were used without further purification.
All the UV-vis spectra were obtained using the SHIMADZU UV-2600i UV-vis spectrophotometer (SHIMADZU, Japan). Fourier-transform infrared spectroscopy from 4,000 to 500 cm−1 was recorded using a Nicolet 6700 FTIR spectrometer (Thermo Inc., USA). All the solutions were prepared through sonication in the ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd., Kunshan, China).
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8

Characterization of Oxidative Reactions

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Ultrapure water was prepared from an ELGA LabWater (ELGA Classic-PURELAB) instrument. Copper(II) chloride dihydrate (analytical grade, ≥ 99%, Sinopharm Chemical Reagent Co., Ltd) and 3, 3′, 5, 5′-Tetramethylbenzidine (analytical grade, ≥ 99%, Macklin Reagent); 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (≥ 98%, OKA); Iron(II) sulfate heptahydrate (analytical grade, ≥ 99%, Sinopharm Chemical Reagent Co., Ltd); 1,2,3-Trihydroxybenzene (analytical grade, Sinopharm Chemical Reagent Co., Ltd); Hydrogen peroxide 30% aqueous solution (in guaranteed grade, Sinopharm Chemical Reagent Co., Ltd); 5,5-Dimethyl-1-Pyrrolidine-N-oxide (≥ 98%, SIAL); The purity of nitrogen and oxygen is greater than 99.999%. These chemicals were used as received without further purification.
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9

Highly Sensitive Enzymatic Biosensor

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All reagents were not purified further before use. Methylbenzene, hydrochloric acid (HCl), lysine (Lys), arginine (Arg), titanium tetrachloride (TiCl4), tetrabutyltitanate, gold (III) chloride (HAuCl4·3H2O), leucine (Leu), cysteine (Cys), proline (Pro), glycine (Gly), barium hydroxide octahydrate (Ba(OH)2·8H2O), formic acid, ethylene diamine tetraacetic disodium (EDTA−2Na), chloroplatinic acid (H2PtCl6), 3,3′,5,5′−tetramethylbenzidine (TMB, ≥99%), L−histidine (L−His), glutathione (GSH), glutamic acid (Glu), and fetal bovine serum were purchased from Macklin Biochemical Co., Ltd. (Shanghai, China). Sodium borohydride (NaBH4), disodium hydrogen phosphate (Na2HPO4), p−Benzoquinone (p−BQ), isopropanol (IPA), potassium nitrate (KNO3), citric acid, ascorbic acid (AA), and sodium nitrate (NaNO3) were purchased from Sinopharm Chemical Reagents Co., Ltd. (Shanghai, China).
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10

Tannic Acid-Copper Complex Colorimetric Assay

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Tannic acid (TA), Cu(NO3)2·3H2O, ethanol, hydrogen peroxide (30 wt%), 3,3′,5,5′-tetramethylbenzidine (TMB), L-proline, glycine, cysteine, alanine, glutamic acid, NaCl, KCl, MnCl2·4H2O, CaCl2, glucose and L-glutathione reduced were purchased from Macklin Biochemical Co., Ltd. Ammonia solution (25–28 wt%) and formaldehyde (37–40 wt%) were purchased from Tianjin Zhiyuan Chemical Co., Ltd. Pluronic® F127 was purchased from Sigma-Aldrich. The qualitative filter paper was purchased from Whatman. All the reagents were used without further purification. Deionized water from a Milli-Q Plus system (Millipore) was used in all experiments.
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