I guess serum-starvation is the simplest option, but for my project is not the best idea. Any other suggestions? Thanks in advance
As far as I know HeLa cells can simply be activated with TNF alpha to induce NFkB signaling (but I didn't do it myself)
Thanks Rodrigo and Christian...just another thing...which concentration? (I know I'm asking a lot...)
I did not understand the point how serum starvation activates NFkB, as you mentioned in your query. IL1 (2-10ng/mL) or TNF (10-20ng/mL) should do it very nicely, with IkB phosphorylation/degradation visible in a few minutes.
Hi Fulvio, you can also try H2O2. I don't know the optimal concentration for your cell line, but usually is around 1 mM. You can check the bibliography. Good luck!
You could activate NF kappaB with pro inflammatory cytokines e.g. TNF or IL-1B
As many have said, TNF or inflammatory cytokines should work.
However, if you don't want to spend hundreds of dollars on lyophilized recombinant TNF, you could use lipopolysaccharide (LPS) to induce inflammatory pathways that activate NF-kB. Some suppliers have phenol-extracted LPS available for under $100 USD.
As mentioned before, NF-kB is activated by a diverse range of stimuli, but one can not use all these drugs randomly rather we have to select drug which best suits our experimental setting. I have been doing NF-kB activation, using serum deprivation, LPS, TPA, H2O2 and Doxo treatments in both adherent and suspension cells. The concentrations of drugs and the kinetics of NF-kB activation is largely cells context dependent. Cycloheximide is also a well know NF-kB activator. I hope very soon you will see my article on this topic........
Hi,
Lipopolysaccharide Induces NF-kB Activation, for more information please refer to:
Achieving Stability of Lipopolysaccharide-Induced NFkB Activation
Markus W. Covert,
Thomas H. Leung,
Jahlionais E. Gaston,
David Baltimore
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