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Kinetic analysis of adenosine triphosphatases (ATPases) by Quench-Flow method (CAT#: STEM-AC-0035-WXH)

Introduction

ATPases are a group of enzymes that catalyze the hydrolysis of a phosphate bond in adenosine triphosphate (ATP) to form adenosine diphosphate (ADP). They harness the energy released from the breakdown of the phosphate bond and utilize it to perform other cellular reactions.




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