Study of cryoenzymology by Quench-Flow method (CAT#: STEM-AC-0043-WXH)

Introduction

A detailed understanding of the mechanism of enzyme action requires a correspondingly detailed knowledge of the structures of the intermediates and transition states on the catalytic pathway, as well as the kinetics and thermodynamics of their interconversion. Cryoenzymology, the use of subzero temperatures and fluid cryosolvents, has the potential to supply this type of information.

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Principle Applications Procedure Materials Notes Related Products

Principle

The principle is to mix the two solutions and then to observe a change in spectroscopic properties of the mixture at different place along the reaction tube during the flow. In the continuous mode of the quenched-flow method, the observation chamber is replaced by a second mixer in which the quenching agent arrives.

Applications

Used to determine fast reaction rates or single turnover rates of enzymatic reactions and to isolate reaction intermediates.

Procedure

Small volumes of solutions are driven through a high efficiency mixer and flow into a delay (or ageing) loop. After a set time, the reaction is stopped (or quenched) by the addition of a chemical quench solution.

Materials

Quench flow

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