Mouse RelA p65 NF-kappa B (NF-kB, Nuclear Factor-kappa B) ELISA Assay Kit. Part No. ELISA-NFKB(m, 96T)

Mouse RelA p65 (Total) ELISA Kit for NF-kB Assays  

Email customersupport@fivephoton.com for current lead time estimate.

(Lead time:  Approximately 2 weeks). 

Fully guaranteed to meet specifications.

• Affinity purified polyclonal capture antibody, with monoclonal HRP-detection antibody.  Sandwich ELISA, strip plate format
  
• Standard peptide concentration:  1800 pg/ml 
• Assay range:  50-1500 pg/ml
• HRP-colorimetric detection at 450 nm
  
• Rapid assay format:  1hr total incubation time at 37^oC  
• Nuclear-Cytoplasmic Isolation kit offered to quantitate the ratio of nuclear relative to cytoplasmic p65.  View available options menu  

 View Protocol and Kit Contents

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Background

The RelA p65 NF-kB (NF-kappa B, NF-{kappa}B) ELISA kit employs the sandwich ELISA(96T) format to quantitate NF-kB p65 protein levels.  The 96-well strip plate is pre-coated with an affinity purified polyclonal capture antibody selective to the RelA (p65) subunit of NF-kB.  After addition of sample, incubation and wash, a monoclonal anti-RelA (p65) HRP-conjugated antibody 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 offered to partition cytoplasmic and nuclear p65.  p65(RelA) in the nuclear and cytoplosmic fractions can be directly measured with this kit.

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Solid Support Sandwich ELISA to RelA p65 NF-kB (NF-kappa B) Methods

Steps of Sandwich ELISA
1.  Equilibrate plate to ambient temperature.

2.  Add sample, incubate 30 min - 1hr, wash. 

3.  Add HRP-Antibody, incubate 30 min -1 hr, wash.

4.  Add color reagents, incubate 15 min.

5.  Add stop solution.  Read Absorbance 450 nm.          

                                                                                                                                                             

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References of RelA p65 NF-kB (NF-kappaB) Assays, Including ELISA

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.
    

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RelA p65 NF-kB (NF-{kappa}B, NF-kappaB, Nuclear Factor-KappaB) ELISA Kit

Safety:  Stop solution contains sulfuric acid.  Avoid all contact and inhalation.
Wear eye protection, gloves.

Storage:  4^oC, 6 months.

Shipping:  Overnight shipping with ice pack is recommended for the NF-kB ELISA Kit.  Also available for international delivery.