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Study of protein phosphorylation by Quench-Flow method (CAT#: STEM-AC-0049-WXH)

Introduction

Protein phosphorylation is a posttranslational modification (PTM) consisting of the addition of phosphate groups to specific amino acid residues on proteins. This PTM has the potential to alter the stability, subcellular location, and enzymatic activity of proteins with diverse roles in cells.




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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