Table 1.

Comparative advantages and disadvantages of biochemistry‐ and imaging‐based approaches to study cellular aspects of ubiquitination

	Advantages	Disadvantages
Biochemistry‐based	Method of choice for substrate identification Chain‐specific Differentiation between mono‐ and polyubiquitination Qualitative/quantitative applications Applicable on tissues and intact organisms	Cell lysis required Cell fractionation often required Quantitative (near) single‐molecule experiments are difficult High‐throughput/high‐content analysis difficult Often time‐consuming Limited spatial resolution Limited temporal resolution Often specialized (and expensive) measurement/analysis set‐ups needed
Imaging‐based	Real‐time live‐cell imaging possible Chain‐specific Native environment No cell fractionation required, use of organelle markers Quantitative single‐molecule imaging possible Qualitative/quantitative applications High‐throughput/high‐content analysis possible Applicable on tissues and intact organisms High spatial resolution High temporal resolution	Not well suitable for substrate identification Potential fixation and permeabilization artefacts Limited differentiation between mono‐ and polyubiquitination Often specialized (and expensive) imaging set‐ups needed