Can anyone tell me if HEK293 cells naturally express the IL10 receptor (either subunit)? Also, has anyone tried a transfection of 293's with a viral vector expressing IL10R?
Hi Sara
293s can be used to express almost anything as they are very amenable to transfection. WHy do you feel you need a viral vector as opposed to simply transfecting the plasmid?
Regarding their endogenous expression of either chain.. may I ask how you were intending to monitor the post virus expression? WHy not use this method?
The reason you should do this yourself is twofold:
1) You should always do things yourself in ordere to see how they look in your own hands;
2) Cell lines vary from lab to lab so one person's reply may not be applicable - did you get your cells directly from ATCC?
Good Luck
G
Thanks for your answer Grant. The reason we are looking to use a viral vector is eventually this experiment will be completed in vivo. Since writing this, we actually did also do a plasmid transfection. We are using the 293s just as a quick preliminary look-see to see if our viral construct is appropriate. Our construct also contains GFP in order that we can monitor GFP expression. All of this is being completed in out own laboratory with our own cells.
Since I wrote this, our virus was working well and we had robust GFP expression. We tried to monitor IL10R1 expression via immunohistochemistry and western blot- both failed due to poor antibodies. Our collaborator has a good antibody for flow, and was able to detected IL10R1 expression in cells through flow.
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