08 August 2019 9 5K Report

Dose anyone have any ideas about the high phosphor-protein level in my western blot basal (negative control)?

Our lab share the same protocol and almost the same regents. Some of the students haven't experienced this problem, but some have. The DMSO in basal is always under 0.1%. We have protease inhibitor in lysis buffer. We tried to always be gentle to the basal sample. Any idea why the basal is so easy to get phosphorylated for some certain protein (for example, its definetly high for pSmad2C but work very well for pAkt, I mean very low for pAkt)? Is phospho-protein samples even not stable in room temperature after denature (denature with sample buffer in 95 degree for 5 min.)?

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