I was wondering if anyone could relate their experience with isolating and analysing protein complexes in complex samples (whole-cell lysates) using mass spectrometry on sucrose gradient fractions.
In particular,
1. does sucrose interfere with the MS run even after TCA precipitation of the proteins (and thereby, hopefully, removal of sucrose)?
2. Are there other gradients (glycerol, PEG etc) that are amenable to subsequent MS analysis of the collected fractions?
Thanks a lot in advance!
For people having the same question, this Nature Protocols paper will help:
https://pubmed.ncbi.nlm.nih.gov/32094787/
I do not know the answer to your main question, but PEG is definitely not compatible with LC/MS.
Check https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7212772/
https://www.sciencedirect.com/science/article/pii/S0005272821000323
Also check https://www.hindawi.com/journals/jamc/2016/8183656/
https://experiments.springernature.com/techniques/sucrose-density-gradient-centrifugation
Also look at this
Chapter System-Wide Profiling of Protein Complexes Via Size Exclusio...
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