Formaldehyde Crosslinking and Its Limitations
Author Information
Author(s): Lars Schmiedeberg, Pete Skene, Aimée Bird, Adrian Deaton, Joshua Z. Rappoport
Primary Institution: Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, United Kingdom
Hypothesis
What is the minimum time required for an interaction to become fixed by formaldehyde crosslinking?
Conclusion
The study establishes that interactions lasting less than 5 seconds are not captured by formaldehyde fixation, revealing significant limitations in the fixation process.
Supporting Evidence
- Interactions lasting less than 5 seconds are invisible after formaldehyde fixation.
- Chromatin immunoprecipitation fails to capture transient interactions.
- The study highlights limitations in the use of formaldehyde as a fixative.
Takeaway
If proteins don't stick around for at least 5 seconds, we can't see them properly under a microscope after using formaldehyde.
Methodology
The study used mutations in the DNA binding protein MeCP2 to analyze the effects of formaldehyde fixation on protein-DNA interactions.
Limitations
The generality of the 5-second threshold for other proteins is not established.
Digital Object Identifier (DOI)
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