Good morning everyone, I have a question regarding some results I obtained. With Western Blot, I observed a high expression of a protein, whereas with confocal microscopy, the same protein shows a significantly reduced expression. In your opinion, what could be the reason for this discrepancy, and what might it depend on?
The discrepancy could stem from differences in detection methods, antibody performance, cellular heterogeneity, subcellular localization, or sample preparation.
You may want to consider:
- Checking antibody performance in both Western Blot and microscopy (e.g., testing different antibodies or using tagged versions of the protein).
- Examining different cell types or subcellular regions in microscopy.
- Reviewing the fixation and imaging protocols.
I hope this gives you some insight into potential causes for the discrepancy
What is your method of observation with confocal microscopy? Immunofluorescence or live-cell microscopy with fluorescent protein labelling?
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