NF-kappa B (NF-KB) - p65 (RelA) Immunofluorescence Labeling Kit (NF-kappaB, NFKB-3)

NF-kappa B (NF-KB) - p65 (RelA)  Immunofluorescence Labeling Kit (NF-kappaB, NFKB-3)
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Price: $395.00
Availability: In Stock
Model: NFKB-3
Manufacturer: Fivephoton Biochemicals

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

Highlights of Kit NFKB-3
  • Offers a visual demonstration of NF-kappa B activation and p65 translocation.
  • Kit provides matched primary and 2o 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.


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.

Methods Overview
Cells are grown on glass cover slips to approximately 85% confluency and treated to activate NF-kappa B.  Cellular proteins are first cross-linked and immobilized with paraformaldehyde (not provided).  The cross-linking reaction is quenched with a provided quenching solution.  Plasma and nuclear membranes are permeabilized with a provided solution that contains a permeabilization reagent and protein blockers.  An included anti-p65 IgG antibody is then applied to cells for 1hr.  Unbound antibody is washed away, and a provided rhodamine tagged Fab fragment secondary antibody is applied for 1 hr.   Unbound secondary antibody is washed away, and a sandwich consisting of slide and cover slip, with antifade solution in-between, is configured.  The antifade solution is allowed to solidify overnight.  The microscope slide-cover slip configuration is now ready for visual and quantitative detection of p65 translocation.

Kit contents
  PBS (10X)
16 ml
  Permeabilization Buffer (10X)
8 ml
  Quenching Buffer (10X)
25 ml
  Intracellular Detection Buffer (10X)
60 ml
  Polyclonal rabbit IgG to p65
120 µl
  Anti-rabbit IgG Rhodamine (Fab fragment)
50 µl
  Forcep set

Pixel Quantitation Software
Related Products:  NF-kappa B Activation Assay Kits
NF-kappa B Activation Assay Kit: click here
p65 Translocation Kit:  click here
p65 total ELISA Kit:  click here

Safety, Storage, Shipping NF-kappa B - p65 Immunofluorescence Kit
Safety:  Irritant.  Avoid ingestion, skin and eye contact
Storage:  Antibodies,  4oC:  Solutions, -20oC
Shipping:  Domestic 2-day Fedex;  $28:  Domestic Overnight Fedex; $42.  International Fedex priority shipping is also available.  Shipped at ambient temperature.  Store as recommended on the label after arrival.  Please inquire with for rate.
kw.  p65 rela translocation assay kit, nf-kappa b activation assay kit, nfkb, translocation assay, immunofluorescence assay.






Category Download Link
Protocol Manual click here
p65 Antibody Sheet
click here
Anti-rabbit IgG-Rhodamine Sheet
click here
MSDS click here