Glutathione S-Transferase Activity Assay

The GST assay reaction mixture contained in 1 ml total volume of 0.1 M K-phosphate buffer, pH 6.5, 1 mM 1-chloro-2,4-dinitrobenzene (CDNB), 1 mM GSH, and enzyme solution. The increase in absorbance was monitored at 340 nm for 3 min at 25 °C. The product extinction coefficient was considered as 9.6 mM⁻¹ cm⁻¹. One GST unit is equivalent to the enzyme amount which conjugates 1 μmole CDNB in 1 min [19 (link)].

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Masoud H.M., Helmy M.S., Darwish D.A, & Ibrahim M.A. (2023). Purification, characterization, and enzyme kinetics of a glutathione S transferase from larvae of the camel tick Hyalomma dromedarii. Journal of Genetic Engineering & Biotechnology, 21, 28.

Publication 2023

Assay Buffer Dinitrobenzene Enzyme Extinction Phosphate

Corresponding Organization : National Research Centre

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Variable analysis

independent variables

Enzyme solution

dependent variables

Increase in absorbance at 340 nm

control variables

0.1 M K-phosphate buffer, pH 6.5
1 mM 1-chloro-2,4-dinitrobenzene (CDNB)
1 mM GSH
Temperature at 25 °C
Reaction time of 3 minutes

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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