Isoproterenol (1-(3,4-dihydroxyphenyl)-2-isopropylaminoethanol hydrochloride, molecular weight: 247.72) and TMQ (2-isopropyl-5-methyl-1, 4-benzoquinone; TMQ) (CAS number 490-91-5, molecular formula C10H12O2, molecular weight 164.20, and purity 99%, Figure 1 ) were procured from Sigma Aldrich (St. Louis, MO, USA). The chemicals and reagents including bovine serum albumin (BSA), 5-sulfosalicylic acid (SSA), naphthylene diamine dihydrochloride, sulphanilamide, phosphoric acid, HEPES, sucrose, 1,4-dithiothreitol (DTT), CHAPS, sodium chloride, protease inhibitors, phenyl methyl sulfonyl fluoride (PMSF), Tween-20, sodium nitrate, 3,3,5,5′-tetramethyl benzidine (TMB), and reduced form of glutathione (GSH) assay kit were purchased from Sigma Aldrich. The enzyme linked immunosorbent assay (ELISA) kit was obtained from R&D Systems, USA. The aspartate transaminase (AST), alanine transaminase (ALT), creatine kinase (CK), and lactate dehydrogenase (LDH) kits were procured form Roche Diagnostics, USA.
Naphthylene diamine dihydrochloride
Manufactured by Merck Group
Sourced in Macao
Naphthylene diamine dihydrochloride is a chemical compound used in various laboratory applications. It serves as a reagent for the detection and quantification of certain substances. The core function of this product is to act as an analytical tool in research and testing environments.
Automatically generated - may contain errors
Lab products found in correlation
Phosphoric acid, by Merck Group (2 mentions)
Sulfanilamide, by Merck Group (2 mentions)
Sulphanilamide, by Merck Group (1 mentions)
Isoproterenol, by Merck Group (1 mentions)
Lactate dehydrogenase ldh, by Roche (1 mentions)
3 3 5 5 tetramethylbenzidine (tmb), by Merck Group (1 mentions)
5 sulfosalicylic acid, by Merck Group (1 mentions)
Enzyme linked immunosorbent assay elisa kit, by R&D Systems (1 mentions)
Creatine kinase, by Roche (1 mentions)
Hepes, by Merck Group (1 mentions)
Sucrose, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Sodium nitrate, by Merck Group (1 mentions)
Protease inhibitor, by Merck Group (1 mentions)
1 4 dithiothreitol dtt, by Merck Group (1 mentions)
Dmem without phenol red, by Thermo Fisher Scientific (1 mentions)
3 protocols using naphthylene diamine dihydrochloride
1
Cardiac Stress Biomarker Evaluation
2
Nitric Oxide Griess Assay Protocol
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A Griess assay was used to determine presence of nitric oxide. In this assay, nitrite, an end product of nitric oxide oxidation, was measured in plasma. The assay reagents included 1% (w/v) sulfanilamide (Sigma-Aldrich Inc., MO) and 0.1% (w/v) naphthylenediamine dihydrochloride (Sigma-Aldrich Inc., MO). Both reagents were dissolved in 2.5% phosphoric acid. Briefly after collecting the plasma, 50 μL of both reagents was added to an equal volume of the plasma in a 96-well round bottom plate. Readings were taken in 5–10 min on an optical density plate reader at 550 nm (Labsystems, Finland).
3
Evaluating Nitric Oxide Levels in LPS-Treated BV2 Cells
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BV2 cells were seeded onto the 96-well plate at 2•5 × 10 4 cells/well and left to attach overnight. Cells were treated with CFS of each CM-LAB at 5-20 % (v/v) in triplicates for 24 h before exposure to 1 µg/ml LPS (Escherichia coli serotype O26:B6; Sigma-Aldrich, St. Louis, MO) in DMEM without phenol red (Invitrogen, Carlsbad, CA) supplemented with 5 % FBS. After incubation for 24 and 48 h in the presence of LPS, nitric oxide (NO) level was determined using the Griess reaction. Culture supernatants (50 µl) of the six CM-LAB were incubated with 50 µl of Griess reagent [1 % sulfanilamide and 0•1 % naphthylene diamine dihydrochloride with 2•5 % phosphoric acid (all Sigma-Aldrich, St. Louis, MO)] for 10 min. The absorbance was read at 530 nm using a microplate reader (MRXll® Dynex Technologies, USA).
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