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

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

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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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.


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ELISA
(54)
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Human
(20)
Mouse
(9)
Rat
(29)
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(1)
Sheep
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Showing Publications 1 - 10 of 75. Show All 75 Publications.
Publications using NBP2-29661 Applications Species
Ma C, Wu C, Jou I et al. PKR activation causes inflammation and MMP-13 secretion in human degenerated articular chondrocytes Thesis 2017 Aug 24 (ELISA, Human) ELISA Human
Miyachi M, Matsuno T, Asano K, Mataga I. Anti-inflammatory effects of astaxanthin in the human gingival keratinocyte line NDUSD-1 Journal of Clinical Biochemistry and Nutrition. 2015 Jan 29 (ELISA, Human)

Details:
RelA/NFkB p65 ELISA kit used for the qualtification of NFkB p65 in nuclear and cytoplasmic fractions (prepared using Nuclear Extraction Kit NBP2-29447 ) from human gingival keratinocytes (NDUSD-1) which were subjected to in-vitro chronic inflammation induced via lipopolysaccharide derived from Escherichia coli O55 followed by treatment or not with anti-inflammatory carotenoid pigment Astaxanthin (Figure 3).
ELISA Human
Zhang F, Ru N, Shang ZH et al. Daidzein ameliorates spinal cord ischemia/reperfusion injury-induced neurological function deficits in Sprague-Dawley rats through PI3K/Akt signaling pathway. Exp Ther Med. 2017 [PMID: 29201192] (ELISA, Rat) ELISA Rat
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
Sun Y, Zhao DL, Liu ZX et al. Beneficial effect of 20-hydroxyecdysone exerted by modulating antioxidants and inflammatory cytokine levels in collagen-induced arthritis: A model for rheumatoid arthritis Mol Med Rep 2017 Aug 29 [PMID: 28901397] (ELISA, Rat) ELISA Rat
Vellai RD, Chandiran S, Pillai SS. GTF-231, a Mixture of Gymnemic Acid, Trigonelline and Ferulic Acid Significantly Ameliorates Oxidative Stress in Experimental Type 2 Diabetes in Rats Can J Diabetes 2017 Jul 17 [PMID: 28728928] (ELISA, Rat) ELISA Rat
Xia R, Ji C, Zhang L. Neuroprotective Effects of Pycnogenol Against Oxygen-Glucose Deprivation/Reoxygenation-Induced Injury in Primary Rat Astrocytes via NF-kB and ERK1/2 MAPK Pathways Cell. Physiol. Biochem. 2017 Jun 27 [PMID: 28662519] (ELISA, Rat) ELISA Rat
Wu Z, Ni J, Liu Y et al. Cathepsin B plays a critical role in inducing Alzheimer's disease-like phenotypes following chronic systemic exposure to lipopolysaccharide from Porphyromonas gingivalis in mice. Brain Behav. Immun. 2017 Jun 10 [PMID: 28610747] (ELISA, Mouse) ELISA Mouse
Chan SH, Hung CH, Shih JY et al. SIRT1 inhibition causes oxidative stress and inflammation in patients with coronary artery disease. Redox Biol. 2017 Jun 02 [PMID: 28601780] (ELISA, Human) ELISA Human
Ma ZG, Xia HQ, Cui SL, Yu J. Attenuation of renal ischemic reperfusion injury by salvianolic acid B via suppressing oxidative stress and inflammation through PI3K/Akt signaling pathway. Braz. J. Med. Biol. Res. 2017 May 15 [PMID: 28513773] (ELISA, Rat) ELISA Rat
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.

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.

The effect of antioxidants and the NFkB p65 pathway in inflammation
NFkB is a transcription factor that plays a role in the expression of genes involved in immune response, inflammation, metastasis, cell survival and more. RelA (p65) is one member of the NFkB mammalian family, alongside other subunits. NFkB subunit...  Read full blog post.

NFkB3-p65: Say that three times fast!
NF-kappa-B is a ubiquitous transcription factor involved in several biological processes such as inflammation, immunity, differentiation, cell growth, tumor genesis and apoptosis. Unlike the majority of transcription factors that reside in the nucleus...  Read full blog post.

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Bioinformatics

Gene Symbol RELA
Entrez