Hello.
I try to detect the presence of cytokines IL-18 and IL-1beta by HEK-blue IL-18 and HEK-blue IL-1beta cells (cells develop by invivogene) these cells are transfected to detect the presence of IL-1b IL-18 in the supernatant. in order to maintain the stability of transfection, these cells are also transfected with resistance genes to selective antibiotics.
the question I ask is when should I add the selection antibiotic? do I have to add them 24 hours before the experiment or when the cells pass? if someone has worked with these cells or with other HEK cells give me a reply please.
thank you in advance.
When it comes to cell lines stably expressing a foreign gene, the media should be supplemented with antibiotics to maintain the specificity. So it is better to add antibiotics in the growth media itself.
This may help you.
https://www.invivogen.com/sites/default/files/invivogen/old/PDF/HEKBlue_IL1b_TDS.pdf
Hello
I worked with the HEKblue IL1b model many years ago.
In my memory, we sellected the cells with antibiotics for one or two weeks. Then we stopped the sellection before our experiments.
The sellection can be done every month or every 2 months.
I apologize for the unprecision of my response as it's been a long time that I used these cells, but I hope it will help you.
Best regards.
@Cédric Rébé
Thank you very much for your answer, what I did is that I added the selection Antibiotic 24h before the experiment. The problem is that when I added the quanti-blue all the wells veered to the blue, even the wells which only contains the hek cells alone. The problem is that the the well contains the culture medium did not turn to the blue what means that the problem does not come from FBS. Do you have an explanation. thank you in advance
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