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How can I induce IFN-gamma production in Jurkat cells using...
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Karl Telemann
How can I induce IFN-gamma production in Jurkat cells using...
maybe this paper can help you. They are using on Jurkat cells PMA (25 ng/ml) and ionomycin (1 uM) for 16h. It seems that as a baseline Jurkats do not produce IFN-g so you have to give them some time for de novo expression before you are able to detect it on the protein level. http://www.nature.com/cr/journal/v18/n9/full/cr2008275a.html good luck with your experiment!
maybe this paper can help you. They are using on Jurkat cells PMA (25 ng/ml) and ionomycin (1 uM) for 16h. It seems that as a baseline Jurkats do not produce IFN-g so you have to give them some time for de novo expression before you are able to detect it on the protein level. http://www.nature.com/cr/journal/v18/n9/full/cr2008275a.html good luck with your experiment!
I am surprised at how long is required to induce IFN-g. In primary T cells, I detect by ICS at 6 hours post-antigen pulse. I'm attempting the Jurkats tonight/tomorrow, and looking for IL2 and IFN-g by ICS. We shall see.... Thanks for the advice.
I am surprised at how long is required to induce IFN-g. In primary T cells, I detect by ICS at 6 hours post-antigen pulse. I'm attempting the Jurkats tonight/tomorrow, and looking for IL2 and IFN-g by ICS. We shall see.... Thanks for the advice.
Thank you for your reply A quick update,: a 24 hour activation with PMA 50 ng/ml + Iono 1uM induced low levels of IFN-g production in my jurkat cells without a massive increase in cell death. Concentrations differed between 15-20 pg/ml and could be measured with my ELISA kit (minimum 9,4 pg/ml). In my own experiments I did not succeed to measure IFN-g in samples activated with 30 or 40 ng/PMA for 24/48 hours, but I dont have a high sensitivity ELISA kit.
Thank you for your reply A quick update,: a 24 hour activation with PMA 50 ng/ml + Iono 1uM induced low levels of IFN-g production in my jurkat cells without a massive increase in cell death. Concentrations differed between 15-20 pg/ml and could be measured with my ELISA kit (minimum 9,4 pg/ml). In my own experiments I did not succeed to measure IFN-g in samples activated with 30 or 40 ng/PMA for 24/48 hours, but I dont have a high sensitivity ELISA kit.
I am trying to use the Jurkat cell line as well, to induce TNFa, or IFNg. I am very surprised by the low induction of those cytokines by Jurkat as compared to primary mononuclear cells. We have compared induction in BSA versus SVF. It appears that SVF does not work at all. BSA gaves us some results, but much less than MNC. We are still working on it. Has someone tested different conditions of stimulation?
I am trying to use the Jurkat cell line as well, to induce TNFa, or IFNg. I am very surprised by the low induction of those cytokines by Jurkat as compared to primary mononuclear cells. We have compared induction in BSA versus SVF. It appears that SVF does not work at all. BSA gaves us some results, but much less than MNC. We are still working on it. Has someone tested different conditions of stimulation?
maybe this paper can help you. They are using on Jurkat cells PMA (25 ng/ml) and ionomycin (1 uM) for 16h. It seems that as a baseline Jurkats do not produce IFN-g so you have to give them some time for de novo expression before you are able to detect it on the protein level.
http://www.nature.com/cr/journal/v18/n9/full/cr2008275a.html
good luck with your experiment!
maybe this paper can help you. They are using on Jurkat cells PMA (25 ng/ml) and ionomycin (1 uM) for 16h. It seems that as a baseline Jurkats do not produce IFN-g so you have to give them some time for de novo expression before you are able to detect it on the protein level.
http://www.nature.com/cr/journal/v18/n9/full/cr2008275a.html
good luck with your experiment!
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I am surprised at how long is required to induce IFN-g. In primary T cells, I detect by ICS at 6 hours post-antigen pulse. I'm attempting the Jurkats tonight/tomorrow, and looking for IL2 and IFN-g by ICS. We shall see.... Thanks for the advice.
I am surprised at how long is required to induce IFN-g. In primary T cells, I detect by ICS at 6 hours post-antigen pulse. I'm attempting the Jurkats tonight/tomorrow, and looking for IL2 and IFN-g by ICS. We shall see.... Thanks for the advice.
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Thank you for your reply
A quick update,: a 24 hour activation with PMA 50 ng/ml + Iono 1uM induced low levels of IFN-g production in my jurkat cells without a massive increase in cell death. Concentrations differed between 15-20 pg/ml and could be measured with my ELISA kit (minimum 9,4 pg/ml). In my own experiments I did not succeed to measure IFN-g in samples activated with 30 or 40 ng/PMA for 24/48 hours, but I dont have a high sensitivity ELISA kit.
Thank you for your reply
A quick update,: a 24 hour activation with PMA 50 ng/ml + Iono 1uM induced low levels of IFN-g production in my jurkat cells without a massive increase in cell death. Concentrations differed between 15-20 pg/ml and could be measured with my ELISA kit (minimum 9,4 pg/ml). In my own experiments I did not succeed to measure IFN-g in samples activated with 30 or 40 ng/PMA for 24/48 hours, but I dont have a high sensitivity ELISA kit.
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jurkat did not secret IFN-γ in my hands either
jurkat did not secret IFN-γ in my hands either
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I am trying to use the Jurkat cell line as well, to induce TNFa, or IFNg. I am very surprised by the low induction of those cytokines by Jurkat as compared to primary mononuclear cells. We have compared induction in BSA versus SVF. It appears that SVF does not work at all. BSA gaves us some results, but much less than MNC. We are still working on it. Has someone tested different conditions of stimulation?
I am trying to use the Jurkat cell line as well, to induce TNFa, or IFNg. I am very surprised by the low induction of those cytokines by Jurkat as compared to primary mononuclear cells. We have compared induction in BSA versus SVF. It appears that SVF does not work at all. BSA gaves us some results, but much less than MNC. We are still working on it. Has someone tested different conditions of stimulation?
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