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Analysis of enzymatic reaction of glutathione transferases by Stopped-flow method (CAT#: STEM-AC-0022-WXH)

Introduction

The glutathione transferases (GSTs; also known as glutathione S-transferases) are major phase II detoxification enzymes found mainly in the cytosol. In addition to their role in catalysing the conjugation of electrophilic substrates to glutathione (GSH), these enzymes also carry out a range of other functions.




Principle

Stopped-flow is an experimental technique for studying chemical reactions with a half time of the order of 1 ms.
In its simplest form, a stopped-flow mixes two solutions. Small volumes of solutions are rapidly and continuously driven into a high-efficiency Ball mixer, so mixing is completed in just a few microseconds. This mixing process then initiates an extremely fast reaction.

Applications

Typically used to gain an understanding of reaction mechanisms, including drug-binding processes or following protein structural changes, stopped-flow spectroscopy enables the study of fast reactions in solution over timescales in the range of millisecond to hundreds of seconds.

Procedure

1. In stopped-flow experiments, two, three, or four sample solutions are rapidly mixed and injected into an observation cell.
2. When the flow is stopped, the kinetics are recorded with a detector best suited to the chemical properties of the solutions and the information of interest (e.g. particle size, the environment of the fluorophore, chromophore).

Materials

Stopped-Flows
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