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LPS-stimulation of Jurkat T-cells to induce TNF-alpha

LPS Jurkat TNF T-cell

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#1 TZ89

TZ89

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Posted 11 June 2013 - 04:29 AM

Hello everybody, I'm trying to get LPS-induced TNF from Jurkat T-cells. I have incubated for 24h with a LPS concentration of 1 - 1000 ng/ml and a cell density of 800.000 cells/ml. But I only detect a maximum TNF-Level of about 50 pg/ml, what is quite too low. I have read that it could has to do with the fact that T-cells have to get activated by PMA/PHA before, but I don't know in what concentrations I have to add these substances. Has any one worked with Jurkat cells before? Or has any one experiences in stimulating them with LPS for inducing cytocines? Thanks a lot! PS: Excuse my bad english, I'm not native





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