I wanted to investigate the inflammatory response of the lipopolysaccharide from the bacterium Porphyromonas gingivalis (P.G.) on human gingival fibroblast cells. To try it out, I wanted to measure the expression level of the proinflammatory cytokine IL-8 with ELISA. As a result of my research, I treated the cells with P.G. lps at different time intervals (1, 3, 6, 9, 12, 24 h) and at a concentration of 0.1, 1, 5, 10 and 50 µg/ml. However, I could not observe a color change according to the standard in any way. What could be the reason for this?
In my case, I cultured gingival fibroblasts till 70% connfluent then stimulate with Pg LPS 1microgram / mL (final concentration) for 24 h. mRNA induction and ELISA results were good. I used ALpha MEM with 10% FBS and 1% p/s
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