It is known that PCSK9 is a kind of secretory protein from hepatocytes and serum PCSK9 is measured by ELISA kit. I was wondering whether it be possible to detect the secreted PCSK9 from cultured HepG2 cells in the medium. Would it be too low to measure? Or anyone being aware of any method?
For my point of view; PCSK9 localized rich in subcellular locations (Nucleus, cytoplasm, ER, Golgi, plasma membrane), and released some extent to extracellular level. But when cells will treated by statins which stimulates SREBP 2 gene for cholesterol synthesis, along with SREBP 2, PCSK9 also raised. For measuring PCSK9 from cell culture: Yes you can do that but you have to measure secreted and purified PCSK9 from culture medium. Also it depends the nature of antibody for PCSK9, conjugated antibodies, detection. I think Chemilumeniscence labeled Ab is much better for sensitivity and linearity.
Thanks for your explanation！ I will give some stimuli and then try to measure PCSK9 in the culture medium.
The detection kits for serum usually works with culture supernatant as well. But PCSK9 is a 74 kDa protein, you can perform western blotting anytime and if you think the concentration will be too low in HepG2 culture supernatant then you can concentrate that supernatant using 3kDa cutoff spin columns before elisa or western.
QconCAT, could be a method to measure your protein of interest.
http://www.polyquant.com/technology/
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