ABTS diammonium salt (AzBTS-(NH4)2)

A radical cation used to assess antioxidant capacity

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货号: ajce51120
CAS: 30931-67-0
别名: 2,2'-联氮双(3-乙基苯并噻唑啉-6-磺酸)二铵盐,AzBTS-(NH4)2
分子式: C18H24N6O6S4
分子量: 548.68
纯度: >98%
溶解度: Water : ≥ 50 mg/mL (91.13 mM)
储存: Store at 2-8°C
库存: 现货

Background

ABTS is a radical cation and a substrate of peroxidases, including horseradish peroxidase (HRP).¹ It has commonly been used to assess antioxidant capacity in the Trolox equivalent antioxidant capacity (TEAC) assay.^2,3 It has a blue color in the presence of sodium persulfate or metmyoglobin but decolorizes upon incubation with antioxidants, and the antioxidant capacity can be determined spectrophotometrically. ABTS has also been used as an enzyme substrate in ELISAs.^4,5 Upon incubation with a peroxidase, it produces a soluble green product that can be quantified by colorimetric detection at 405 nm.¹

1.Bhattarai, H.D., Paudel, B., Lee, H.S., et al.Antioxidant activity of Sanionia uncinata, a polar moss species from King George Island, AntarcticaPhytother. Res.22(12)1635-1639(2008) 2.Huang, D., Ou, B., and Prior, R.L.The chemistry behind antioxidant capacity assaysJ. Agric. Food Chem.53(6)1841-1856(2005) 3.Walker, R.B., and Everette, J.D.Comparative reaction rates of various antioxidants with ABTS radical cationJ. Agric. Food Chem.57(4)1156-1161(2009) 4.Reyna-Bello, A., Eleizalde, M.C., and Silva, A.M.Assessment of chromogen suitability in ELISA for the detection of anaplasmosis and trypanosomosisJ. Immunoassay Immunochem.28(1)1-11(2007) 5.Su, Y.C., and Wong, A.C.Detection of staphylococcal enterotoxin H by an enzyme-linked immunosorbent assayJ. Food Prot.59(3)327-330(1996)

Protocol

Kinase experiment:

Enzyme activity is determined photometrically using a temperature controlled multi-mode plate reader or alternatively in a UV/Vis spectrophotometer. Reactions are initiated by addition of the enzyme. Enzyme activity is measured over a period of 10 min at 25°C at the appropriate wavelength for the substrate. One unit (1 U) is defined as the amount of enzyme that converts 1 μmol substrate per minute. Various H2O2 concentrations (0-1250 μM, enzyme concentrations (0.27-54 nM) and substrate concentrations are used to determine the enzyme activity. The activity of rPsaDyP vs ABTS is determined in 100 mM sodium acetate buffer at pH 3.8 and a final H2O2 concentration of 125 μM. Production of the ABTS cation radical is studied at 420 nm (ε420 36,000 L mol-1 cm-1)[2].

参考文献:

[1]. Matsuda H, et al. Evaluation of ELISA with ABTS, 2-2'-azino-di-(3-ethylbenzthiazoline sulfonic acid), as the substrate of peroxidase and its application to the diagnosis of schistosomiasis. Jpn J Exp Med. 1984 Jun;54(3):131-8.
[2]. Lauber C, et al. Identification, heterologous expression and characterization of a dye-decolorizing peroxidase of Pleurotus sapidus. AMB Express. 2017 Aug 23;7(1):164.