Nov 12, 2023
In vitro dephosphorylation
- 1University of California, San Diego
Protocol Citation: Leonardo A Parra-Rivas 2023. In vitro dephosphorylation. protocols.io https://dx.doi.org/10.17504/protocols.io.kqdg3xb1pg25/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: November 12, 2023
Last Modified: November 12, 2023
Protocol Integer ID: 90820
Abstract
In vitro dephosphorylation
100 μg of α-syn GST fusion protein on beads were resuspended in alkaline
phosphatase (ALP) buffer supplemented with 250 units of ALP (Thermo Scientific
Cat# EF0651).
The mixes were incubated for 60 min or 180 min at 37 °C with
orbital shaking (Eppendorf ThermoMixer F1.5).
Samples were then analyzed by
NuPAGE and immunoblotted or alternatively used in pull-down experiments with
brain lysates.