Human RelA (p65) NF-kappa B (NF-kB, Nuclear Factor-kappa B) ELISA Assay Kit (Part No. NFKB-ELISA(h))

Human RelA(p65)Total NF-kappaB ELISA Kit; (96T)

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

(Lead time: Approximately 2 weeks).

Detects total human p65(RELA)
Affinity purified polyclonal capture antibody, with polyclonal HRP-conjugated detection antibody
Standard Peptide Concentration: 2250 pg/ml
Assay Range: 100 - 2000 pg/ml
HRP-Colorimetric detection at 450nM
Strip plate format
Rapid assay: 1 hr total incubation time at 37^oC (i.e. with plate and HRP-detection Ab).
Nuclear-Cytoplasm Isolation Kit offered as an option to separately quantitate nuclear and cytoplasmic p65 as a method to estimate levels of NF-kappa B activation
Separate p65 ELISA kits are offered for human, mouse and rat

Number of Kits	Cost (USD)
1	$635.00
2	$1150
Nuclear-Cytoplasm Isolation Kit	$150

View Protocol and Kit Contents

Use the protocol shipped with the kit for your experiment.

Icon Image: P65-p50 dimer; Source PDB.

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 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 second polyclonal 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 to provide an estimate of NF-kappa B activation by calculation of the nuclear p65 relative to cytoplasmic p65 concentration ratio.

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.

Product Citation:

Anti-RelA siRNA-Encapsulated Flexible Liposome with Tight Junction-Opening Peptide as a Non-invasive Topical Therapeutic for Atopic Dermatitis. 2019;42(7):1216-1225. doi: 10.1248/bpb.b19-00259.

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

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.
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.
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.
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.
Jin, S. et al. (2005) A simplified probe preparation for ELISA-based NF-kappaB activity assay. J Biochem Biophys Methods 65 (1), 20-29.
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.
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.
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.
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.
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.
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.
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.
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.

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 domestic shipping with ice pack is recommended for the NF-kB ELISA Kit (Cost: $52.00).

Category	Download Link
Protocol Manual	click here
MSDS	click here

NF-KAPPA B ACTIVATION ASSAY KIT \| NFKB, NF-kB, NF-{kappa}B, Nuclear Factor-Kappa B \| Part NFKB-2 NFKB-2 $459.00	Antibody to p50 subunit of NF-kappa B (P50Ab) P50Ab $287.95	Mouse RelA p65 NF-kappa B (NF-kB, Nuclear Factor-kappa B) ELISA Assay Kit. Part No. ELISA-NFKB(m, 96T) ELISA-NFKB(m) $635.00	NF-KAPPA B (NF-KB) - p65 (RelA) IMMUNOFLUORESCENCE LABELING KIT (NF-kappaB, NFKB-3) NFKB-3 $450.00
HUMAN AMYLOID-BETA 1-40 100pg-3000ng/ml Assay Range ELISA KIT (Beta-Amyloid, AB, ABeta). Part No. hAB1-40ELISA (kw. beta amyloid, amyloid beta) hAB1-40ELISA $780.00	Rat RelA p65 NF-kappa B (NF-kB, Nuclear Factor-kappa B) ELISA Assay Kit (Part No. NFKB-ELISA (r, 96T)) NFKB-ELISA (r, 96T) $635.00

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