Hi, I have run a couple of Western blots for tyrosine hydroxylase in mammalian SH-SY5Y cells and have noticed 2 distinct bands rather than just one, as is shown in most of the literature. Some other papers do show 2 bands, but don't really address it. I thought it could be a phosphorylated form of the protein, but the jump in size looks a little big for that?
Interestingly, the higher band only appears in the second and third lanes where the lysate loaded was from cells I have put through a differentiation protocol; the first lane is from undifferentiated cells.
Does anybody have any ideas about how to interpret this/have experience in blotting for TH and could offer any advice? Thank you!
Hi Naomi Thorne
* Please check the antibody's literature, most of them show the image of the blot in their website and you can always put the catalogue number and search the publications using the same antibody.
* Single band is observed in TH procured from CST, Abcam, Novus and Thermo.
Thanks
If we have to exclude problems related primary AB and other technical issues, you would make sure about the source of these bands by knocking down or out the gene and detect again for the presence of these strange bands.
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