I am taking 2*106 cells, centrifuge them and remove the media. I add fresh media, which have already the concentration of LPS as 2ug/ml. Then I culture these cell for given time
THP-1 cell with DMSO pre-treament can also induce IL-12 expression. BTW, did any control group show a positive result in your assay ? LPS should be worked.....
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1906042/
2ug/mL seems very high, is that normal for THP-1 cells? I typically use 1-100ng/mL for my cell cultures. Have you checked that you did not kill all your cells? Can you measure IL-12 in the culture supernatant?
I repeated assay assay with 1ug/ml LPS and assay design was as follows: I induced 2*106 cells with 1.2 % DMSO and than induced with 1ug/ml of LPS for 0,4,12 and 24 hrs. But no expression of IL-12. And I can,nt measure IL-12 in supernatant.
I have same probllem with you. I add LPS to treat THP1 macrophage but no increasing of IL-1beta and IL-12. Have you solved the problem?
I could not proceed that project, but you cn check the life of THP-1 cell if these r too old and had multiplied in many generations, 2nd perform the flowcytometry to cnfirm that weither these r THP-1 real? any way gud luck with your project and do share with me if u get the expression
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