Hi all,
I have just used mouse brain samples to investigate levels of GAD65 via western blotting (ab239372). I got very specific staining, but the band appeared to be in a different weight around 50kDa. Does anyone know possible reasons as to why this happened? Is this unspecific binding of the antibody? Or improper handling of the sample?
Thanks!
Run positive and negative controls and dilute the Antibody concentration
Hello,
Besides what you mentioned, you can also try to analyze this in a different way altogether.
Could this protein of 50kDa be a truncated form of GAD65?
For your reference I have attached two research articles. You may get some help from these articles.
https://doi.org/10.1007/s11064-005-6772-0
https://doi.org/10.1371/journal.pone.0102056
Best.
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