Hello, I'm having a bit of trouble doing the Western blot experiment.
Even though I load the same sample, the band appears inconsistently (in all the trials).
(I usually store the samples at -20°C and boil them at 95°C for 5 min before loading.)
This sample is harvested from bacteria-infected mammalian cells and I extracted only the mammalian cell protein by RIPA-buffer lysis method.
I wonder if anyone has an experience like this and how to solve this problem.
Thank you so much in advance!
Inconsistent band patterns in Western blotting can result from various factors, including issues with sample preparation, electrophoresis, transfer, blocking, antibody incubation, washing steps, detection sensitivity, sample heterogeneity, and experimental reproducibility. To address these issues and achieve more consistent results:
By carefully controlling each step of the Western blotting process and addressing potential sources of variability, you can enhance the consistency and reliability of your results. Additionally, including appropriate controls and performing replicate experiments can help validate your findings.
- Badges
- Science topic
- Similar topics
- Agricultural Science
- Horticulture