The cationic surfactant tested was benzethonium chloride (BEC) (IUPAC name: benzyl-dimethyl-[2-[2-[4-(2,4,4-trimethylpentan-2-yl) phenoxy] ethoxy] ethyl] azanium chloride; commercial name: Hyamine 1622). It was purchased from Fluka Chemie GmbH (Buchs, Switzerland), having a purity higher than 96%, CAS no. 121-54-0 and molecular weight of 448.08 g/mol. Chemical reagents for surfactant detection (chloroform, disulfide blue VN 150, ethanol, 1 N sulfuric acid), and biochemical and histological reagents were obtained from Sigma-Aldrich (St. Louis, MO, USA), having a purity higher than 98%.
1 n sulfuric acid
Manufactured by Merck Group
Sourced in Germany
1 N sulfuric acid is a common laboratory reagent used for various analytical and experimental purposes. It has a concentration of 1 normal (1 N), which indicates the molarity of the sulfuric acid solution. This product is frequently utilized in titration procedures, pH adjustment, and other laboratory applications requiring a standardized sulfuric acid solution.
Automatically generated - may contain errors
Lab products found in correlation
Ethanol, by Merck Group (1 mentions)
Chloroform, by Merck Group (1 mentions)
Spectramax plus 384 microplate reader, by Molecular Devices (1 mentions)
Tetramethylbenzidine solution, by Merck Group (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Trypsin edta, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
Nystatin, by Merck Group (1 mentions)
Sodium hypochlorite, by Merck Group (1 mentions)
Phalloidin and dapi, by Merck Group (1 mentions)
Sodium tungstate, by Merck Group (1 mentions)
Disodium hydrogen orthophosphate, by Merck Group (1 mentions)
Sodium nitroprusside, by Merck Group (1 mentions)
96 well half area microplate, by Corning (1 mentions)
Tmb substrate solution, by Merck Group (1 mentions)
Horseradish peroxidase hrp conjugated anti m13 antibody, by Sino Biological (1 mentions)
Spectramax 190 microplate reader, by Molecular Devices (1 mentions)
5 protocols using 1 n sulfuric acid
1
Benzethonium Chloride Surfactant Characterization
2
Quantification of Serum SPARC Levels
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Sandwich ELISA (Human SPARC ELISA kit; #LS-F12653; LifeSpan BioSciences, Inc., Seattle, WA, USA) was used to measure the levels of SPARC in different serum samples. Serum was diluted with enzyme immunoassay (EIA) buffer (containing 1% bovine serum albumin and 0.05% Tween 20 in phosphate buffer) from the ELISA kit and incubated for 2 h at 37°C. After 4 washes with EIA buffer, mouse anti-human IgG horseradish peroxidase-conjugated antibodies (1:2,500 dilution) from the ELISA kit were added and incubated for 30 min at 4°C. After a further 4 washes, 100 µl tetramethylbenzidine solution (Sigma-Aldrich, St. Louis, MO, USA) was added and incubated for 30 min at room temperature. The reaction was subsequently stopped with 100 µl 1 N sulfuric acid (Sigma-Aldrich). The experimental procedure was conducted according to the instruction manual and the results were measured using a SpectraMax® Plus 384 Microplate Reader (Molecular Devices, LLC, Sunnyvale, CA, USA) at 450 nm.
3
Bioactivity Evaluation of Terminalia chebula
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T. chebula fruits (voucher no. TERCHE 0413 deposited at plant herbarium YNUH, Republic of Korea) were collected from HNB Garhwal University (a central university) campus, Srinagar, Garhwal, India. Organic solvents like ethyl acetate, acetone, and methanol and reagents such as 2,2-Diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl (DPPH), Dulbecco's modified Eagle's medium (DMEM), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), glutaraldehyde (50%), trypsin-EDTA, fetal bovine serum, penicillin-streptomycin, nystatin, phalloidin and DAPI, sodium nitroprusside, sodium hydroxide, disodium hydrogen orthophosphate, sodium hypochlorite, 1N sulfuric acid, and sodium tungstate were purchased from Sigma Aldrich (Yongin, South Korea). Monoclonal anticytokeratin and TRITC labeled secondary-IgG antibody was obtained from Sigma Aldrich (St. Louis, U.S.). Live/Dead viability/cytotoxicity Kit was purchased from Abcam (Cambridge, U.K.). Keratinocytes cell was obtained from American Type Culture Collection (ATCC) (Manassas, VA, USA) and Fibroblasts cell line (L929) was procured from Korean Cell Bank (Seoul, South Korea).
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T. chebula fruits (voucher no. TERCHE 0413 deposited at plant herbarium YNUH, Republic of Korea) were collected from HNB Garhwal University (a central university) campus, Srinagar, Garhwal, India. Organic solvents like ethyl acetate, acetone, and methanol and reagents such as 2,2-Diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl (DPPH), Dulbecco's modified Eagle's medium (DMEM), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), glutaraldehyde (50%), trypsin-EDTA, fetal bovine serum, penicillin-streptomycin, nystatin, phalloidin and DAPI, sodium nitroprusside, sodium hydroxide, disodium hydrogen orthophosphate, sodium hypochlorite, 1N sulfuric acid, and sodium tungstate were purchased from Sigma Aldrich (Yongin, South Korea). Monoclonal anticytokeratin and TRITC labeled secondary-IgG antibody was obtained from Sigma Aldrich (St. Louis, U.S.). Live/Dead viability/cytotoxicity Kit was purchased from Abcam (Cambridge, U.K.). Keratinocytes cell was obtained from American Type Culture Collection (ATCC) (Manassas, VA, USA) and Fibroblasts cell line (L929) was procured from Korean Cell Bank (Seoul, South Korea).
4
Quantification of Mouse Serum Ig Isotypes
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Total IgG and IgG2a and IgG1 isotypes were measured in mouse serum, by indirect ELISA, using 96-well plates. In brief, 100 μl of PBS at concentration of 10 g/ml at pH = 7.4, was added to each well (1 g/well) and incubated overnight at 4°C. Then, it was washed 3 times with Phosphate Buffered Saline with Tween (PBS-T). Mouse sera were diluted 1:100 and 100 μl was added to the wells and incubated for 2 h at 37°C. After washing, 100 μl of peroxidase-conjugated antibody with 1:100000 dilutions were added to each well and incubated for 90 minutes at 37°C. Following few washing steps, 100 μl of chromogenic substrate (TMB/H2O2, Razi-institute, Iran) was added to each well, the enzyme activity was stopped, using 1N sulfuric acid (Merck-Germany), and the absorbance was read at 450 nm.
5
Monoclonal Phage ELISA for YKL-40 Detection
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A monoclonal phage ELISA was performed after three rounds of panning. Several 96-Well Half-Area Microplates (Corning, New York, NY, USA) were coated overnight at 4 °C, with 30 μL per well of 1 μg/mL hYKL-40, and each well was blocked with 5% skim milk in PBS for 1 h at RT. The amplified phages of individual clones from the third round of panning were added and incubated for 1 h at 37 °C. After washing four times with PBS-T, horseradish peroxidase (HRP)-conjugated anti-M13 antibody (1:5000, Sino Biological, Beijing, China) was incubated for 1 h at 37 °C. After washing it four times with PBS-T, a TMB substrate solution (Sigma-Aldrich, St. Louis, MO, USA) was added for 8 min, and the reaction was stopped with 1 N sulfuric acid (Merck, Darmstadt, Germany). The absorbance was measured at 450 nm using a SpectraMax 190 Microplate Reader (Molecular Devices, Sunnydale, CA, USA).
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