In the N-glycomics experiment I am unable to see any glycans nor any protein. I am doubting whether it could be the separation step of protein and N-glycans might be the problem.
Actual experiment: I used methanol and centrifuged it and collect the supernatant for N-glycans analysis.
Doubt: I checked that methanol should be stored at 15 -25 degree, however it was kept around 4 degree for long period of time. Could this be the reason for methanol also precipitates out the glycans along with proteins.
Check https://www.sciencedirect.com/science/article/pii/S0008621519301673
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757665/
Also check https://microbialcellfactories.biomedcentral.com/articles/10.1186/s12934-020-1280-0
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5877797/
Higher yields than acetic acid when the derivatization was performed at 37 °C for 16 h. When higher temperatures and very acidic conditions are applied, partial degradation of the glycans will be induced with loss of sialic acids.
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