NF-KAPPA B (NF-KB) - p65 (RelA) IMMUNOFLUORESCENCE LABELING KIT (NF-kappaB, NFKB-3)

NF-kappa B - p65 (NF-kB, Nuclear Factor-kappa B, RelA) Immunofluorescence Translocation Labeling Kit

• Offers a visual demonstration of NF-kappa B activation and p65 translocation.
• Kit provides matched primary and 2^o fluorophore-tagged (rhodamine, Fab fragment) antibodies that detect p65 in nucleus and cytoplasm.
• Applicable for most mammalian cells (e.g. human, mouse, rat, hamster, canine, rabbit, equine).
• NF-kB activation is quantified by measuring immunofluorescence intensity of p65 using pixel quantification software.*
• Sufficient solutions to label p65 in 40 microscope slides.

     Protocol

Background

The NF-kB complex resides in the cytoplasm in its inactive state consisting of the inhibitory IkB subunit assembled with the p65-p50 heterodimer.  Upon stimulus, the IkB inhibitory subunit is ubiquitinated and degraded, exposing nuclear localization signals on the p65 (RelA) subunit, which then translocates into the nucleus.  The antibodies provided with this kit are employed to label p65 in nucleus and cytoplasm.  Fluorescent microscopy and pixel quantitation software are then employed to estimate the emission intensity of p65.