In vitro binding assay by Microscale Thermophoresis (MST)

MZ Marita Zoma
LC Laura Curti
DS Dheeraj Shinde
DA Domenico Albino
AM Abhishek Mitra
JS Jacopo Sgrignani
SM Sarah N. Mapelli
GS Giada Sandrini
GC Gianluca Civenni
JM Jessica Merulla
GC Giovanna Chiorino
PK Paolo Kunderfranco
AC Alessia Cacciatore
AK Aleksandra Kokanovic
AR Andrea Rinaldi
AC Andrea Cavalli
CC Carlo V. Catapano
GC Giuseppina M. Carbone
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EZH2 recombinant protein His-tagged (Epigentek, E24031-1) was labeled with Monolith His-Tag Labeling Kit (MO-L008). ERG recombinant protein (Origene, TP308093) at a concentration ranging from 0.4uM to 2.44E-05uM or BSA (0.02uM), was incubated with labeled EZH2 (5 nM fixed concentration) at room temperature for 5 min. Microscale thermophoresis measurements were performed utilizing a Monolith NT.115 instrument (NanoTemper Technologies GmbH, Munchen, Germany). The measurements were performed using Premium capillaries at 30% LED excitation and high MST power.

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