Hello,
I am planning to enrich O-GlucNAc proteins in Jurkat T-cells by metabolic labeling before performing drug-treatment with Cisplatin.
I have some practical questions regarding this:
1. Is it absolute necessary to have the cells cultured for at least 6 passages before performing drug-treatment?
2. Do I need to have the cells being serum starved before drug-treatment? Since cells will be treated with Cisplatin in 16 hours to induce apoptosis. Starving the cells before hand could induce unnecessary stress to the cells, hence affecting the results?
3. I will need to culture cells with medium containing GlucNAz to metabolically label glycosylation proteins for at least 72 hours. Will starving the cells after this could also induce unwanted changes on the glycosylation profile?
Thank you so much
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