I was using K-562 cell line as a negative control for my Western-blot analysis of EAAT2. It turned out to be positive with a nice band. I was using a primary monoclonal mouse antibody sc-365634 (Santa-Cruz) which had nice reviews.
I have found two studies discussing the role of glutamate receptors in Meg-01 and K-562 cells and also the expression of SLC1A3 gene.
Nonethless, the proteinatlas does not show any hint that K-562 would express EAAT2.
https://www.proteinatlas.org/ENSG00000110436-SLC1A2
What do you think? Is it the fault of the antibody or the cell line is truly positive?
Alexandr Zarivnij
In fact, I don't use antibody produced by santacruz although it has the good reviews (I don't know the reason but I have heard the fail cases from my colleague). So, you'd better to validate western blotting by using other antibody from major company (abcam, cell signaling, merck...).
As I review protein atlas and the reference you attached, I think you'd better to change cell line for negative control. I didn't clearly review the reference. But, as you know, RNA level does not completely convert protein. In protein atlas, we can find section of pathogen. According to this data, you can find other cell line by negative control (very low expression of EAAT2). You need to trust data of protein atlas because they are specialist for antibody.
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