I want to determine the constitutive NF-kappaB in MM cell lines by western or/and RT-PCR but I don't know the criteria. Are negative/positive control cell lines needed to see which is constitutively activated?
if you mean constitutive expression of NF-kappaB then you should demonstrate the mRNA / protein levels in your cell lines (using qPCR/western blots). I don't think the negative control is necessary for that if you are using a commercial antibody for the western and run a 'water' or no RT for the qPCR.
If you mean constitutive activity then you might want to look at IKappaB or the kinase. I believe that is the negative regulator. phospho-specific antibodies with or without inhibs could prove as useful controls for western blots.
(I assume that your treatment/stimulation/activation induces NFkB activation)
Extract proteins from treated and untreated cells. Perform a western blot using an antibody targeting phosphorylated NFkB. Stripe your membrane and stain again using an anti-NFkB antibody. Finally, check loading by actin staining.
NFkB bands should be similar, showing the constitutive expression, but you should see stronger pNFkB signal in the treated cells.
Cell Signalling has well working NFkB antibodies, as well as TNFa stimulated and resting cell lysates to be used as controls for the NFkB phosphorylation.
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