The results for activation I have been observing have been scattered and inconsistent. I am looking for reagents or a protocol to reliably induce NLRP3 inflammasome activation and show a clear difference to the control. I am also concerned that the RAW 264.7 cells are reacting in an adverse manner to something I am overlooking. Any tips on how to keep RAW 264.7 cells from activating prematurely would be greatly appreciated.
Alexander Soloniuk I'm working with primary cells neutrophils and monocytes and I stimulated my cells with LPS (3 hrs) and ATP (30 min), this is my positive control for NLRP3 inflammasome. So, I think you need to check if your cells can express CD14, because some types of cells didn't express this molecule and in this case its not possible stimulate with LPS.
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