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NF- kappa B - p65 Total ELISA Kit (Human, 96T; Part hNFKBp65-ELISA, kw NFKB ELISA, p65, NF-kappa B, Assay Kit)

NF- kappa B - p65 Total ELISA Kit (Human, 96T; Part hNFKBp65-ELISA, kw NFKB ELISA, p65, NF-kappa B, Assay Kit)
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Price: $645.00
Availability: In Stock
Model: hNFKBp65-ELISA

DetailsManuals & Downloads
Human NF-kappa B (p65)Total ELISA Kits
Sandwich ELISA format (96T)
 
Highlights
  • Complete kit in sandwich ELISA format providing convenience and high sensitivity. 
  • Strip plate configuration allowing separate use and storage of sections of the ELISA plate
  • Rapid assay format.  1 incubation for 60 min
  • Wide assay Range:  50 pg/ml - 15 ng/nl.  Sensitivity:  24 pg/ml
  • Can be adapted to p65 translocation assays by using an available Nuclear-Cytoplasmic protein Isolation kit to isolate p65 (Part NPI-1)
Lead time approximately 2 weeks.
 
 
Price (USD)
Number of Kits
Number of Assays
645
1
96T


 
Specifications
Species
Standard Peptide
Assay Range
Human
16 ng/ml
50 pg/ml - 15 ng/ml
                               


Background
The Human p65 NF-kB (NF-kappa B, NF-{kappa}B) ELISA kit employs the sandwich ELISA(96T) format to quantitate total NF-kB p65 protein levels.  The 96-well strip plate is coated with an affinity purified monoclonal capture antibody selective to the RelA (p65) subunit of NF-kB.  After addition of sample, incubation and wash, a second monoclonal anti-RelA (p65) HRP-conjugated antibody labelled with biotin is added to each well.  Color reagents are added to generate a colorimetric reaction, which corresponds to the concentration of RelA (p65) in the sample.
 
A Nuclear-Cytoplasmic fractionation kit is available to partition cytoplasmic and nuclear p65 (Part NPI-1).  p65(RelA) in the nuclear and cytoplosmic fractions can be directly measured with this ELISA kit to provide an estimate of NF-kappa B activation by calculating the relative nuclear p65 to cytoplasmic p65 concentration.

Methods Overview:
 
The included ELISA micro-strip plate is provided pre-coated with an a monoclonal capture antibody selective to the antigen.  Samples are pre-mixed with a biotin-linked detection antibody and streptavidin-HRP, which are then added to the plate.  The plate is washed after a 60 min incubation period.  Color reagents are added, resulting in a reaction with HRP that yields a blue colored solution.  A stop solution is applied to terminate the reaction, and the solution is measured at 450nm.  
 
   Solid Support Sandwich ELISA to RelA p65 NF-kB (NF-kappa B) Methods                                                                                                                               

ELISA Assay Steps
  1. Equilibrate plate to ambient temperature.
  2. Add sample, wash.
  3. Add HRP-Antibody, incubate 1hr at 37oC, wash.
  4. Add color reagents A and B, incubate 10 min at 37oC.
  5. Add stop solution.  Read Absorbance 450 nm.
 
References on NF-kappa B
  1. Baudy, A.R. et al. (2009) A robust in vitro screening assay to identify NF-kappaB inhibitors for inflammatory muscle diseases. Int Immunopharmacol 9 (10), 1209-1214.
  2. Dann, S.G. et al. (2004) Chromatin immunoprecipitation assay on the rainbow trout opsin proximal promoters illustrates binding of NF-kappaB and c-jun to the SWS1 promoter in the retina. Exp Eye Res 78 (5), 1015-1024.
  3. Hassan, N.J. et al. (2009) Development of an insect-cell-based assay for detection of kinase inhibition using NF-kappaB-inducing kinase as a paradigm. Biochem J 419 (1), 65-73.
  4. Hellweg, C.E. et al. (2003) Generation of stably transfected Mammalian cell lines as fluorescent screening assay for NF-kappaB activation-dependent gene expression. J Biomol Screen 8 (5), 511-521.
  5. Jin, S. et al. (2005) A simplified probe preparation for ELISA-based NF-kappaB activity assay. J Biochem Biophys Methods 65 (1), 20-29.
  6. Kaur, J. et al. (2008) Chemopreventive activity of lantadenes on two-stage carcinogenesis model in Swiss albino mice: AP-1 (c-jun), NFkappaB (p65) and P53 expression by ELISA and immunohistochemical localization. Mol Cell Biochem 314 (1-2), 1-8.
  7. Klinke, D.J., 2nd et al. (2008) Modulating temporal control of NF-kappaB activation: implications for therapeutic and assay selection. Biophys J 94 (11), 4249-4259.
  8. Li, J.L. et al. (2007) An optimized assay for transcription factor NF-kappaB with dsDNA-coupled microplate. Colloids Surf B Biointerfaces 55 (1), 31-37.
  9. Moon, K.Y. et al. (2001) A cell-based assay system for monitoring NF-kappaB activity in human HaCat transfectant cells. Anal Biochem 292 (1), 17-21.
  10. Noursadeghi, M. et al. (2008) Quantitative imaging assay for NF-kappaB nuclear translocation in primary human macrophages. J Immunol Methods 329 (1-2), 194-200.
  11. Sun, C.S. et al. (2008) Anti-sense morpholino oligonucleotide assay shows critical involvement for NF-kappaB activation in the production of Wnt-1 protein by HepG2 cells: oncology implications. J Biomed Sci 15 (5), 633-643.
  12. Tomita, K. et al. (1999) Functional assay of NF-kappaB translocation into nuclei by laser scanning cytometry: inhibitory effect by dexamethasone or theophylline. Naunyn Schmiedebergs Arch Pharmacol 359 (4), 249-255.
  13. Wang, J. et al. (2006) Exonuclease-mediated ELISA-like assay for detecting DNA-binding activity of transcription factors: measurement of activated NF-kappaB. Biotechniques 41 (1), 79-88, 90.
Safety, Storage, Shipping, NFKB-p65 ELISA Kit
Safety:  Stop solution contains acid.   Avoid ingestion, skin and eye contact.
Storage:  2 - 8oC.  Expires 6 months after manufacture.

Shipping.  2-day or Overnight delivery recommended.  International delivery is also available

kw.  NFKB, p65, RelA, ELISA, Assay Kit, NF-kappa B


 

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