There was also a similar pre-print in early October, where Maria Hondele's group showed that tagging can have a huge impact on the phase-separation behavior of proteins: https://doi.org/10.1101/2024.10.04.616694
That reminds me of this great paper where everything starts out by realizing that the GFP tagged transcription factor behaved differently, leading to find new epigenetic regulations mechanisms that we obliterated by the tag https://pubmed.ncbi.nlm.nih.gov/22579476/
It’s key to remind about “side effects”of fluorescent tags.
We recently found that, in an ex vivo leukocytes recruitment assay, labeling cells with antibodies conjugated with different dyes (PE vs AF488 vs BV421) had different outcomes.
Some dyes were more prone to stick to the endothelium !!!
Comments
https://rupress.org/jcb/article/163/2/257/33516/Formation-of-stacked-ER-cisternae-by-low-affinity
Corollary: localization of That under Those Conditions is merely where That is under Those Conditions.
FPs are victims of their own success. You can see how messed up they get, if you bother to look. Granted, many people don't.
Most experiments do not allow this insight -Western Blot, proteomics, drug screens, most structural biology etc.
And no experiment is silent.
https://bsky.app/profile/lindorfflarsen.bsky.social/post/3l5twzlq44e2w
We recently found that, in an ex vivo leukocytes recruitment assay, labeling cells with antibodies conjugated with different dyes (PE vs AF488 vs BV421) had different outcomes.
Some dyes were more prone to stick to the endothelium !!!