I need to do a phenotypic characterisation of human IPSC-derived hepatocyte-like cells by flow cytometry.
I've used HepG2 but it didn't seem to work well for albumin antibody.
The percentage of albumin-positive cells was the same for both positive(HepG2) and negative controls(human IPSC).
Shoul I try a different antibody?
Came across this article that may help. They incubate the cells in LDL A488 then measure it via flow.
Perhaps this is better than a specific antibody. Albumin can stick to a lot of things which could explain the signal in both groups
http://www.nature.com/mt/journal/v20/n1/full/mt2011234a.html
You may consider the heparg cells , have a look at www.heparg.com,
Dear Sofia,
Perhaps some of these lines could be useful for you:
http://www.lgcstandards-atcc.org/Products/All/PTA-5565.aspx?slp=1&geo_country=es
http://www.upcyte.com/products/human-upcyte-hepatocytes.html
HepG2 has always been a reliable positive control for albumin in our hands. As for the HepaRG suggestion, you would need to keep in mind that depending on your culture conditions not all cells will be positive for albumin, see slide attached.
True, there are two cell populations in a Heparg monolayer (but only hepatocytes on a spheroid mode of culture). The good thing is that it is albumin and not alpha foeto protein like on the hepg2. And you can consider cyp3a4 staining too that will be very intense (same level as in the primary heps) whereas you will see nothing in the hepg2. Look at actin staining too around the bile canaliculi (simple protocol that i will be pleased to forward) , the Heparg form functional bile canaliculi ...good luck if you try with the hepg2. Christophe
Dear Christophe, good point about the cyp3a4, I will give it a go. I am very interested in that protocol you mentioned too, if you could forward it.