Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric)

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ELISA: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) [NBP2-29661] - Standard curve using the internal RelA standard.
ELISA: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) [NBP2-29661] - Nuclear translocation of NF-kB p65 with LPS in Jurkat cells. Jurkat cells, grown to a density of 2X10(6) cells/ml, were treated with LPS (15 ...read more
ELISA: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) [NBP2-29661] - Immunofluorescence indicates NF-kB p65 subunit was expressed exclusively in AA-G groups and AST-G cytoplasm. High glucose stimulation impact ...read more

Product Details

Summary
Reactivity Hu, Mu, RtSpecies Glossary
Applications ELISA
Suitable Sample Type
Cell and tissue lysates
Standard Curve Range
15.6 - 1000 ng/ml
Sensitivity
15.6 ng/ml

Order Details

Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) Summary

Description
APPROPRIATE CONTROLS TO INCLUDE: Following is a list of suggested controls to include with each analysis: 1. No capture antibody added to well. 2. No lysate added to well. 3. No capture antibody or lysate added to well. 4. Positive control: use a cell line or tissue known to constitutively express p65 or a recombinantly expressed p65. 5. Negative control: use a cell line or tissue known to not express p65.
Specificity
For the detection of cytoplasmic, nuclear and total free NF-kB/p65 in the nucleus of either cells or tissues.
Standard Curve Range
15.6 - 1000 ng/ml
Sensitivity
15.6 ng/ml
Assay Type
Sandwich ELISA
Sample Volume
0.1 ml
Kit Type
ELISA Kit (Colorimetric)
Gene
RELA

Applications/Dilutions

Application Notes
Please note that this kit has not yet been tested or used with any of the reporter cell lines and is therefore not guaranteed for detection of the target protein these samples. Contains reagents and protocol to prepare whole, nuclear, and cytoplasmic cell fractions. Multiple samples can be analyzed in a low-volume, high-throughput format. Full analysis complete in just hours. Allows direct measurement of changes in p65 translocation. Allows study of NF-kB activation without gel-shift assay. [This product is the same as Imgenex's NF-kappaB p65 ActivELISA Kit, IMK-503].
Publications
Read Publications using
NBP2-29661 in the following applications:

Reactivity Notes

Human, mouse, rat and sheep reported in the literature.

Packaging, Storage & Formulations

Storage
Storage is content dependent.
Preservative
0.02% Sodium Azide

Kit Components

Components
  1. Recombinant p65 Standard 2 vials lyophilized(0.42 ug/vial)
  2. 100X Protease Inhibitor Cocktail (PIC) 100 ul
  3. 10X Hypotonic Lysis Buffer 10 ml
  4. 10X PBS 2 x 50 ml
  5. 100 mM PMSF 10 ml
  6. 10 mM DTT (for Nuclear Lysis Buffer) 500 ul
  7. 10% Detergent Solution 10 ml
  8. 1X Nuclear Extraction Buffer 10 ml
  9. 1 M DTT (for Nuclear Extraction from tissue) 100 ul
  10. AKP-Conjugated Secondary Ab 10 ul
  11. BSA 2 x 0.5 g
  12. Capture Antibody 200ul
  13. Coating Buffer 2 x 10 ml
  14. ELISA Plates 2
  15. Detecting Antibody 200 ul
  16. pNPP 4 x 5 mg
  17. pNPP Substrate Buffer 2 x 10 ml
  18. Manual 1

Notes

Additional items required for the ELISA (not included NF-kB/p65 ActivELISATM Module): Distilled water 96-well ELISA plate reader

Alternate Names for Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric)

  • MGC131774
  • nf kb elisa kit
  • nf kb p65
  • nfkb elisa kit
  • NFkB p65
  • NF-kB p65
  • NFKB3
  • NFKB3v-rel avian reticuloendotheliosis viral oncogene homolog A (nuclear factor ofkappa light polypeptide gene enhancer in B-cells 3 (p65))
  • Nuclear factor NF-kappa-B p65 subunit
  • Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3transcription factor p65
  • p65 elisa kit
  • p65
  • p65RelA
  • rela p65
  • RelA
  • v-rel reticuloendotheliosis viral oncogene homolog A (avian)
  • v-rel reticuloendotheliosis viral oncogene homolog A, nuclear factor of kappalight polypeptide gene enhancer in B-cells 3, p65

Background

Activation of the NF-kB pathway can be triggered by many factors including TNFalpha, UV, IL-1, lipopolysaccharide (LPS), mitogens, and phorbol esters. NF-kB is controlled by a family of inhibitory proteins called, IkBs. IkB proteins are phosphorylated by IkB kinase complex consisting of at least three proteins, IKK1/alpha, IKK2/Beta, and IKK3/gamma. External stimuli such as tumor necrosis factor or other cytokines initiate a signal transduction cascade that leads to the activation of IkB-kinase complex, which then specifically phosphorylates IkBalpha on Serine-32 and Serine-36. Phosphorylation of these sites leads to ubiquitination of IkBalpha and subsequent degradation by the 26 S proteasome. Degradation of IkBalpha results in unmasking of the nuclear localization signal of NF-kB dimers, which subsequently translocates to the nucleus and acts as a transcription factor for genes controlling inflammatory cytokines, adhesion molecules, and other proteins. Thus the nuclear levels of p65 may correlate positively with the activation of NF-kB pathway.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. ELISA Kits are guaranteed for 6 months from date of receipt.

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Supplier Logo

Publications for RelA/NFkB p65 ELISA Kit (NBP2-29661)(75)

We have publications tested in 5 confirmed species: Human, Mouse, Rat, Mouse , Sheep.

We have publications tested in 1 application: ELISA.


Filter By Application
ELISA
(54)
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Filter By Species
Human
(20)
Mouse
(9)
Rat
(29)
Mouse
(1)
Sheep
(1)
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Showing Publications 1 - 10 of 75. Show All 75 Publications.
Publications using NBP2-29661 Applications Species
Hara K, Hamada C, Wakabayashi K et al. Scavenging of reactive oxygen species by astaxanthin inhibits epithelial-mesenchymal transition in high glucose-stimulated mesothelial cells PLoS ONE 2017 Sep 19 [PMID: 28926603] (ELISA) ELISA
Yamada K, Morinaga Y, Yanagihara K et al. Azithromycin inhibits MUC5AC induction via multidrug-resistant Acinetobacter baumannii in human airway epithelial cells. Pulm Pharmacol Ther. 2014 Jun 06 [PMID: 24910464] (ELISA, Human)

Details:
Fig 6: NCI-H292 human airway epithelial cells. The p65 subunit of NF-kB was measured in nuclear extracts from cells stimulated with the pathogen multidrug-resistant Acinetobacter baumannii (MDRAB), and then left untreated or treated with azithromycin (AZM) or clarithromycin (CAM). MDRAB had a time-dependent effect on NF-kB p65 translocation. AZM but not CAM supressed the effects of MDRAB in this model system.
ELISA Human
Wang K, Xiao J, Peng B et al. Retinal structure and function preservation by polysaccharides of wolfberry in a mouse model of retinal degeneration. Sci Rep. 2014 Dec 23 (ELISA, Mouse)

Details:
RelA/NFkB p65 / NF-kB/p65 ActivELISA kit used for measurement of NF-kB p65 in the retina of WT and wolfberry polysaccharides- and PBS-treated rd10 mice by ELISA application (Figure 6C).
ELISA Mouse
Wihastuti TA, Sargowo D, Agoes A et al. Level of Circulating Endothelial Cells and Expression of Nuclear Factor Kappa Beta of Human's Peripheral Blood Mononuclear Cells in Subjects with Certain Conditions. Acta Med Indones 2014 Jan 01 [PMID: 24760806] (ELISA, Human)

Details:
PBMC, Table I. The authors detailed their lysis and extraction buffer recipes as well as analysis strategy of NF-kB translocation in the Methods section of the manuscript.
ELISA Human
Li R, Zang A, Zhang L et al. Chrysin ameliorates diabetes-associated cognitive deficits in Wistar rats. Neurol. Sci. 2014 Apr 16 [PMID: 24737349] (ELISA, Rat)

Details:
Rat brain, Fig 4.
ELISA Rat
Hirsch J, Chalkley RJ, Bentley T et al. Double impact of cigarette smoke and mechanical ventilation on the alveolar epithelial type II cell. Crit Care 2014 Mar 25 [PMID: 24666941] (ELISA, Rat)

Details:
Nuclear fractions of primary rat avelolar type II cells, Table 1.
ELISA Rat
Singh DP, Chopra K. et al. Flavocoxid, dual inhibitor of cyclooxygenase-2 and 5-lipoxygenase, exhibits neuroprotection in rat model of ischaemic stroke. Pharmacol. Biochem. Behav. 2014 Feb 18 [PMID: 24561313] (ELISA, Rat)

Details:
Rat brain, Fig 4.
ELISA Rat
Lin X, Zhang S, Huang R et al. Protective effect of tormentic acid from Potentilla chinensis against lipopolysaccharide/d-galactosamine induced fulminant hepatic failure in mice. Int. Immunopharmacol. 2014 Feb 21 [PMID: 24560903] (ELISA, Mouse)

Details:
Mouse liver, Fig 4F.
ELISA Mouse
Yang ZH, Liu LB, Zhao LN et al. Permeability of the blood-tumor barrier is enhanced by combining vascular endothelial growth factor with papaverine. J. Neurosci. Res. 2014 Feb 12 [PMID: 24523141] (ELISA, Rat)

Details:
Rat brain microvessel fractions, Fig 7C.
ELISA Rat
Huang WC, Tsai PJ, Huang YL et al. PGE2 production is suppressed by chemically-synthesized delta-7-eicosatrienoic acid in macrophages through the competitive inhibition of COX-2. Food Chem. Toxicol. 2014 Jan 27 [PMID: 24480038] (ELISA, Mouse)

Details:
Stimulated Raw264.7 cells, Fig 5A.
ELISA Mouse
Show All 75 Publications.

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Product General Protocols

View specific protocols for RelA/NFkB p65 ELISA Kit (NBP2-29661): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for RelA/NFkB p65 ELISA Kit (NBP2-29661). (Showing 1 - 2 of 2 FAQs).

  1. Can we perform this ELISA on the frozen brain tissues or every time you need fresh tissue?
    • We have not evaluated this kit on brain tissue, neither fresh nor frozen. The the kit is quality controlled using the recombinant protein standard in the kit. However, the kit has been published using different types of tissues. Please follow this link for representative publications, including brain tissue NBP2-29661. Ultimately, the researcher will need to empirically determine if the kit is suitable for their brain tissue and how the results from frozen and fresh brain tissue compare.
  2. How do I thaw the 100X protease inhibitor cocktail that comes with the NF-kB p65 ELISA kit?
    • It is recommended that you leave it at room temp for 30mins, or 4 hours at 4C 

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Blogs on RelA/NFkB p65. Showing 1-10 of 147 blog posts - Show all blog posts.

Killing two birds with one stone: Treating inflammation and cancer by inhibiting prolyl-4-hydroxylase-1
By Jamshed Arslan Pharm.D. The cell’s oxygen-sensing machinery comprises prolyl-4-hydroxylases (P4Hs 1-3, PHDs 1-3, or EGLN 1-3) and their canonical target hypoxia-inducible factors (HIFs). When oxygen levels...  Read full blog post.

Killing two birds with one stone: Treating inflammation and cancer by inhibiting prolyl-4-hydroxylase-1
By Jamshed Arslan Pharm.D. The cell’s oxygen-sensing machinery comprises prolyl-4-hydroxylases (P4Hs 1-3, PHDs 1-3, or EGLN 1-3) and their canonical target hypoxia-inducible factors (HIFs). When oxygen levels...  Read full blog post.

Killing two birds with one stone: Treating inflammation and cancer by inhibiting prolyl-4-hydroxylase-1
By Jamshed Arslan Pharm.D. The cell’s oxygen-sensing machinery comprises prolyl-4-hydroxylases (P4Hs 1-3, PHDs 1-3, or EGLN 1-3) and their canonical target hypoxia-inducible factors (HIFs). When oxygen levels...  Read full blog post.

Killing two birds with one stone: Treating inflammation and cancer by inhibiting prolyl-4-hydroxylase-1
By Jamshed Arslan Pharm.D. The cell’s oxygen-sensing machinery comprises prolyl-4-hydroxylases (P4Hs 1-3, PHDs 1-3, or EGLN 1-3) and their canonical target hypoxia-inducible factors (HIFs). When oxygen levels...  Read full blog post.

Killing two birds with one stone: Treating inflammation and cancer by inhibiting prolyl-4-hydroxylase-1
By Jamshed Arslan Pharm.D. The cell’s oxygen-sensing machinery comprises prolyl-4-hydroxylases (P4Hs 1-3, PHDs 1-3, or EGLN 1-3) and their canonical target hypoxia-inducible factors (HIFs). When oxygen levels...  Read full blog post.

Killing two birds with one stone: Treating inflammation and cancer by inhibiting prolyl-4-hydroxylase-1
By Jamshed Arslan Pharm.D. The cell’s oxygen-sensing machinery comprises prolyl-4-hydroxylases (P4Hs 1-3, PHDs 1-3, or EGLN 1-3) and their canonical target hypoxia-inducible factors (HIFs). When oxygen levels...  Read full blog post.

Killing two birds with one stone: Treating inflammation and cancer by inhibiting prolyl-4-hydroxylase-1
By Jamshed Arslan Pharm.D. The cell’s oxygen-sensing machinery comprises prolyl-4-hydroxylases (P4Hs 1-3, PHDs 1-3, or EGLN 1-3) and their canonical target hypoxia-inducible factors (HIFs). When oxygen levels...  Read full blog post.

Killing two birds with one stone: Treating inflammation and cancer by inhibiting prolyl-4-hydroxylase-1
By Jamshed Arslan Pharm.D. The cell’s oxygen-sensing machinery comprises prolyl-4-hydroxylases (P4Hs 1-3, PHDs 1-3, or EGLN 1-3) and their canonical target hypoxia-inducible factors (HIFs). When oxygen levels...  Read full blog post.

Killing two birds with one stone: Treating inflammation and cancer by inhibiting prolyl-4-hydroxylase-1
By Jamshed Arslan Pharm.D. The cell’s oxygen-sensing machinery comprises prolyl-4-hydroxylases (P4Hs 1-3, PHDs 1-3, or EGLN 1-3) and their canonical target hypoxia-inducible factors (HIFs). When oxygen levels...  Read full blog post.

Killing two birds with one stone: Treating inflammation and cancer by inhibiting prolyl-4-hydroxylase-1
By Jamshed Arslan Pharm.D. The cell’s oxygen-sensing machinery comprises prolyl-4-hydroxylases (P4Hs 1-3, PHDs 1-3, or EGLN 1-3) and their canonical target hypoxia-inducible factors (HIFs). When oxygen levels...  Read full blog post.

Showing 1-10 of 147 blog posts - Show all blog posts.

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Bioinformatics

Gene Symbol RELA
Entrez