Journal
INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY
Volume 135, Issue -, Pages -Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.biocel.2021.105978
Keywords
Protein complexes; Calibration standards; Oligomerization; Quantitative microscopy
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Funding
- Centre of Membrane Protein Research (COMPARE, University of Birmingham, UK)
- Centre of Membrane Protein Research (COMPARE, University of Nottingham, UK)
- DFG (Deutsche Forschungsgemeinschaft, Germany) [HE4559/6-1]
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Quantitative assessment of protein complexes, particularly in the context of cellular signaling, has become a pressing objective in cell biology. Recent advancements in single molecule fluorescence microscopy have led to different approaches for counting protein copy numbers in various cellular structures. Novel calibration protocols are needed to address challenges in molecular counting experiments.
Quantitative assessment of protein complexes, such as receptor clusters in the context of cellular signalling, has become a pressing objective in cell biology. The advancements in the field of single molecule fluorescence microscopy have led to different approaches for counting protein copy numbers in various cellular structures. This has resulted in an increasing interest in robust calibration protocols addressing photophysical properties of fluorescent labels and the effect of labelling efficiencies. Here, we want to give an update on recent methods for protein counting with a focus on novel calibration protocols. In this context, we discuss different types of calibration samples and identify some of the challenges arising in molecular counting experiments. Some recently published applications offer potential approaches to tackle these challenges.
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