Hi all, so I did western blot to detect GAPDH as my control. Despite repeating three times, all my western blot results shows variability in different aspects.
I've tried to use commercial blocking buffer instead of milk to reduce noise, I also filter my TBS-T with 0.22 um filter. In addition, I tried loading the same sample with increasing concentration, but it shows less intense band instead of the normal ones. I also tried to carefully remove the bubble from my PVDF membrane, and carefully prepare the sample loaded by multiple check while pipetting. Could this be a problem with my laboratory techniques? I would really appreaciate it if you have any suggestions. Thank you.
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