Microscale thermophoresis quantifies biomolecular interactions under previously challenging conditions

► Microscale thermophoresis (MST) quantifies protein-protein binding in cell lysate. ► The KD of Grb2-homodimerization is determined. ► Competitive binding of small molecules and peptides to proteins is analyzed. ► GPCR membrane proteins are characterized using label-free MST. ► Syt1-binding to PIP2-liposomes and the cooperativity of Ca2+/PIP2 and syt1 is quantified.