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.

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

Name: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric)
Brand: Novus Biologicals
SKU: NBP2-29661
Price: 679 USD

(95 Publications)

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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

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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. 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].
Specificity	For the detection of cytoplasmic, nuclear and total free NF-kB/p65 in the nucleus of either cells or tissues. Human, mouse, rat and sheep reported in the literature.
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

Dilutions

ELISA

Publications

Read Publications using
NBP2-29661 in the following applications:

ELISA
70 publications

Packaging, Storage & Formulations

Storage

Storage is content dependent.

Kit Components

Components

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

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)(95)

We have publications tested in 4 confirmed species: Human, Mouse, Rat, Sheep.

We have publications tested in 1 application: ELISA.

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ELISA

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(22)

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Showing Publications 1 - 10 of 95. Show All 95 Publications.

Publications using NBP2-29661	Applications	Species
Ahmed M. Awad, Sally L. Elshaer, Rajashekhar Gangaraju, Rania R. Abdelaziz, Manar A. Nader Ameliorative effect of montelukast against STZ induced diabetic nephropathy: targeting HMGB1, TLR4, NF-kappa B, NLRP3 inflammasome, and autophagy pathways Inflammopharmacology 2023-07-27 [PMID: 37498374]
ARAI M, TSUJI M, TSUCHIYA H. Protective Effects of Fucoidan Against Interleukin-1b-induced Inflammation in SW982 Human Synovial Cells. The Showa University Journal of Medical Sciences. 2014-09-27 (ELISA, Human) Details: RelA/NFkB p65 ELISA Kit used for measuring NF-kB-free p65 in the nuclear fractions of SW982 cells (human synovial sarcoma cell line) which were stimulated with IL1 beta for 24 hours with or without 1 hour Fucoidan pre-treatment (Fig 4).	ELISA	Human
Srinivas BK, Bourdi A, O'Regan JD et al. Interleukin-1? Disruption Protects Male Mice From Heart Failure With Preserved Ejection Fraction Pathogenesis Journal of the American Heart Association 2023-06-22 [PMID: 37345828] (ELISA)	ELISA
Lu Y, Xing C, Zhang C et al. Promotion of IL?17/NF?k B signaling in autoimmune thyroid diseases Experimental and Therapeutic Medicine 2022-12-06 [PMID: 36588813] (ELISA) Details: Serum Samples	ELISA
Lu Y, Xing C, Zhang C et al. Promotion of IL?17/NF?k B signaling in autoimmune thyroid diseases Experimental and Therapeutic Medicine 2022-12-06 [PMID: 36588813] (ELISA) Details: Serum Samples	ELISA
Younis NS, Ghanim AMH The Protective Role of Celastrol in Renal Ischemia-Reperfusion Injury by Activating Nrf2/HO-1, PI3K/AKT Signaling Pathways, Modulating NF-kappa b Signaling Pathways, and Inhibiting ERK Phosphorylation Cell biochemistry and biophysics 2022-03-01 [PMID: 35157199] (ELISA)	ELISA
Abdel-Latif RT, Wadie W, Abdel-Mottaleb Y Et al. Reposition of the anti-inflammatory drug diacerein in an in-vivo colorectal cancer model Saudi Pharm J 2022-02-11 [PMID: 35145347] (ELISA) Details: Citation using the HRP version of this antibody.	ELISA
Uniyal A, Kotiyal A, Gadepalli A et al. Epigallocatechin-3-gallate Improves Chronic Alcohol Induced Cognitive Dysfunction in Rats by Interfering with the Neuro-inflammatory, Cell Death and Oxido-nitrosative Stress Pathways Metab Brain Dis 2021-08-13 [PMID: 34386880] (ELISA)	ELISA
Joharapurkar A, Patel V, Kshirsagar S et al. Effect of dual PPAR-alpha/gamma agonist saroglitazar on diabetic retinopathy and oxygen-induced retinopathy European journal of pharmacology 2021-03-19 [PMID: 33753107] (ELISA, Rat)	ELISA	Rat
Liu Y, Shen W, Chen Q et al. Inhibition of RAGE by FPS-ZM1 alleviates renal injury in spontaneously hypertensive rats European Journal of Pharmacology 2020-06-01 [PMID: 32502492] (ELISA, Rat)	ELISA	Rat
Chen YP, Chen CT, Liu TP et al. Catcher in the rel: Nanoparticles-antibody conjugate as NF-kappa B nuclear translocation blocker Biomaterials 2020-01-01 [PMID: 32247937] (ELISA)	ELISA
Liu Z, Zhang H, Wang H et al. Magnolol Alleviates IL-1beta-Induced Dysfunction of Chondrocytes Through Repression of SIRT1/AMPK/PGC-1alpha Signaling Pathway J. Interferon Cytokine Res. 2020-01-09 [PMID: 31916911]
Godugu C, Pasari LP, Khurana A et al. Crocin, an active constituent of Crocus sativus ameliorates cerulein induced pancreatic inflammation and oxidative stress Phytother Res 2019-11-25 [PMID: 31769107] (ELISA)	ELISA
He L, Zhao X, He L LINC01140 Alleviates the Oxidized Low-Density Lipoprotein-Induced Inflammatory Response in Macrophages via Suppressing miR-23b Inflammation 2019-11-20 [PMID: 31748847] (ELISA, Human)	ELISA	Human
Fouad AA, Abdel-Aziz AM, Hamouda AAH Diacerein Downregulates NLRP3/Caspase-1/IL-1 beta and IL-6/STAT3 Pathways of Inflammation and Apoptosis in a Rat Model of Cadmium Testicular Toxicity Biol Trace Elem Res 2019-08-10 [PMID: 31401744] (ELISA)	ELISA
Khurana A, Anchi P, Allawadhi P et al. Superoxide dismutase mimetic nanoceria restrains cerulein induced acute pancreatitis Nanomedicine (Lond) 2019-07-01 [PMID: 31267840] (ELISA, Mouse)	ELISA	Mouse
He LP, Zhao XS, He LP. Abnormally expressed miR-23b in Chinese Mongolian at high cardiovascular risk may contribute to monocyte/macrophage inflammatory reaction in atherosclerosis.. Biosci. Rep. 2018-10-12 [PMID: 30314997] (ELISA, Human)	ELISA	Human
Tekula S, khurana A, Anchi P, Godugu C. Withaferin-A attenuates multiple low doses of Streptozotocin (MLD-STZ) induced type 1 diabetes. Biomedicine & Pharmacotherapy. 2018-10-01 [PMID: 30119216] (ELISA, Mouse)	ELISA	Mouse
Rojas E, Zhang C, Bowman MAH. Enzyme-modified non-oxidized LDL (ELDL) induces human coronary artery smooth muscle cell transformation to a migratory and osteoblast-like phenotype. Ann Arbor [PMID: 30097618] (ELISA, Human)	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. 2017-01-01 [PMID: 29201192] (ELISA, Rat)	ELISA	Rat
Epigallocatechin gallate supplementation protects against renal injury induced by fluoride intoxication in rats: Role of Nrf2/HO-1 signaling. Thangapandiyan S, Miltonprabu S, et al. Toxicol Rep [PMID: 28962222] (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 2017-09-19 [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 2017-08-29 [PMID: 28901397] (ELISA, Rat)	ELISA	Rat
Ma C, Wu C, Jou I et al. PKR activation causes inflammation and MMP-13 secretion in human degenerated articular chondrocytes Redox Biol 2017-09-05 [PMID: 28869834] (ELISA)	ELISA
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 2017-07-17 [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. 2017-06-27 [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. 2017-06-10 [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. 2017-06-02 [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. 2017-05-15 [PMID: 28513773] (ELISA, Rat)	ELISA	Rat
Kang HJ, Chung DH, Sung CO et al. SHP2 is induced by the HBx-NF-kB pathway and contributes to fibrosis during human early hepatocellular carcinoma development. Oncotarget 2017-04-18 [PMID: 28460481] (ELISA, Human)	ELISA	Human
Stano A, Leaman DP, Kim AS et al. Dense array of spikes on HIV-1 virion particles. J. Virol. 2017-04-26 [PMID: 28446665] (ELISA)	ELISA
Liu Y, Xiong Y, Xing F et al. Precise Regulation of miR-210 Is Critical for the Cellular Homeostasis Maintenance and Transplantation Efficacy Enhancement of Mesenchymal Stem Cells in Acute Liver Failure Therapy Cell Transplant. 2017-05-09 [PMID: 27983913] (ELISA, Human)	ELISA	Human
Huang Q, Bai F, Nie J et al. Didymin ameliorates hepatic injury through inhibition of MAPK and NF-kB pathways by up-regulating RKIP expression. Int. Immunopharmacol. 2017-01-01 [PMID: 27912149] (ELISA, Mouse)	ELISA	Mouse
Tsai KL, Hung CH, Chan SH et al. Baicalein protects against oxLDL-caused oxidative stress and inflammation by modulation of AMPK-alpha. Oncotarget. 2016-11-08 [PMID: 27776344] (ELISA, Human)	ELISA	Human
Anti-inflammatory effects of astaxanthin in the human gingival keratinocyte line NDUSD-1 Miyachi M, Matsuno T, Asano K J Clin Biochem Nutr [PMID: 26060346] (ELISA, Human)	ELISA	Human
Sachdeva AK, Chopra K. Lycopene abrogates Abeta(1-42)-mediated neuroinflammatory cascade in an experimental model of Alzheimer's disease J. Nutr. Biochem. 2015-03-20 [PMID: 25869595] (ELISA, Rat)	ELISA	Rat
Fouad A, Amin E, Ahmed A Naringenin and Vanillin Mitigate Cadmium-Induced Pancreatic Injury in Rats via Inhibition of JNK and p38 MAPK Pathways PJ 2020-06-18 (Rat)		Rat
Rajasekar N, Nath C, Hanif K Shukla R. Intranasal Insulin Administration Ameliorates Streptozotocin (ICV)-Induced Insulin Receptor Dysfunction, Neuroinflammation, Amyloidogenesis, and Memory Impairment in Rats. Mol. Neurobiol. 2016-10-11 [PMID: 27730514] (ELISA, Rat)	ELISA	Rat
Fouad AA, Qutub HO, Al-Melhim WN. Punicalagin alleviates hepatotoxicity in rats challenged with cyclophosphamide. Environ. Toxicol. Pharmacol. 2016-06-01 [PMID: 27310207] (ELISA, Rat)	ELISA	Rat
Chan SH, Chu PM, Kao CL et al. Oleic acid activates MMPs up-regulation through SIRT1/PPAR-gamma inhibition: a probable linkage between obesity and coronary arterial disease. J. Biochem. 2016-04-12 [PMID: 27072559] (ELISA, Human)	ELISA	Human
Rajasekar N, Nath C, Hanif K, Shukla R. Inhibitory Effect of Memantine on Streptozotocin-Induced Insulin Receptor Dysfunction, Neuroinflammation, Amyloidogenesis, and Neurotrophic Factor Decline in Astrocytes. Mol. Neurobiol. 2015-12-11 [PMID: 26660109] (ELISA, Rat) Details: RelA/NFkB p65 ELISA Kit was used for ELISA analysis of nuclear fractions from Rat astrocytoma cell line (C6) which was incubated with 100 uM Streptozotocin in the absence or presence of memantine and memantine alone (Fig 9a).	ELISA	Rat
Dwivedi S, Rajasekar N, Hanif K et al. Sulforaphane Ameliorates Okadaic Acid-Induced Memory Impairment in Rats by Activating the Nrf2/HO-1 Antioxidant Pathway. Mol. Neurobiol. 2015-10-03 [PMID: 26433376] (ELISA, Rat) Details: RelA/NFkB p65 ELISA Kit was used for estimation of NF-kB-free p65 in the nuclear fraction of cerebral cortex and hippocampus regions from rats subjected to treatments with sulforaphane and/or okadaic acid (neurotoxin).	ELISA	Rat
Hung CH, Chan SH, Chu PM, Tsai KL et al. Quercetin is a potent anti-atherosclerotic compound by activation of SIRT1 signaling under oxLDL stimulation Mol Nutr Food Res. 2015-07-23 [PMID: 26202455] (ELISA, Human) Details: RelA/NFkB p65 ELISA kit (Imgenex IMK-503) was used for quantitative ELISA of nuclear NFkB on HUVECs that were exposed to quercetin followed by oxidized low-density lipoprotein/oxLDL-mediated activation of NK-kB. Moreover, HUVEC pretreatment with DCHC, AICAR, SC79, DPI and SNP (NO donor) was found to result in a significant effect on inhibiting NK-kB activation, while, EX-527, Compound C, LY294002 (AKT inhibitor) and eNOS siRNA antagonized the protective effect of quercetin (Figure 7A).	ELISA	Human
Vors C, Pineau G, Drai J et al. Postprandial endotoxemia linked with chylomicrons and LPS handling in obese vs lean men: a lipid dose-effect trial J. Clin. Endocrinol. Metab. 2015-07-07 [PMID: 26151336] (ELISA, Human)	ELISA	Human
Huang WC, Tsai TH, Huang CJ et al. Inhibitory effects of wild bitter melon leaf extract on Propionibacterium acnes-induced skin inflammation in mice and cytokine production in vitro Food Funct. 2015-06-22 [PMID: 26098998] Details: NF-kappaB p65 ActivELISA Kit was used for the determination of NF-kB activation in THP-1 cells, human monocytic THP-1 cells (3 × 10^6 cells per mL) that were treated with total phenolic extract of wild bitter melon, luteolin and/or Propionibacterium acnes (M.O.I. = 75). See the full text for experimental detail and the data is shown in Fig 5.
Yu S, Liu LB, Cao Y et al. Combining Bone Marrow Stromal Cells with Green Tea Polyphenols Attenuates the Blood-Spinal Cord Barrier Permeability in Rats with Compression Spinal Cord Injury. J. Mol. Neurosci. 2015-05-26 [PMID: 26007330] (ELISA, Rat)	ELISA	Rat

Xu P, Cai F, Liu X, Guo L. Sesamin inhibits lipopolysaccharide-induced proliferation and invasion through the p38-MAPK and NF-kB signaling pathways in prostate cancer cells Oncol. Rep. 2015-03-31 [PMID: 25845399] (ELISA, Human)	ELISA	Human
Lakshmi A, Subramanian SP. Tangeretin ameliorates oxidative stress in the renal tissues of rats with experimental breast cancer induced by 7,12-dimethylbenz[a]anthracene Toxicol. Lett. 2014-09-02 [PMID: 24995432] (ELISA, Rat) Details: NFkB p65 ELISA Kit / NF-kB/p65 ActivELISA-TM kit used for quantification of nuclear levels of free p65 in kidney tissue homogenate of Rats that were subjected to experimental breast cancer induction with 7,12-dimethylbenz[a]anthracene/DMBA followed by treatment with Tangeretin, a citrus polymethoxyflavone (Fig 4).	ELISA	Rat
Ghaith K, Shalaby MA, Ramadan A Rosuvastatin Restrains the Headway of Experimentally Induced Liver Fibrosis: Involvement of NF-?B and Nrf2/HO-1 Signaling Pathway International Journal of Veterinary Science 2022-11-26 (ELISA)	ELISA
Shi Z, Lian A, Zhang F. Nuclear factor-kB activation inhibitor attenuates ischemia reperfusion injury and inhibits Hmgb1 expression. Inflamm. Res. 2014-09-11 [PMID: 25209109] (ELISA, Mouse) Details: NFkB p65 ELISA kit used for measuring p65 (sub unit of NF-kB) translocation into the nucleus in experiments involving cardiac ischemia reperfusion injury in a transplantation model treated or not with nuclear factor-kB activation inhibitor dehydroxymethylepoxyquinomicin/DHMEQ (Fig. 4)	ELISA	Mouse
Rajasekar N, Dwivedi S, Nath C et al. Protection of streptozotocin induced insulin receptor dysfunction, neuroinflammation and amyloidogenesis in astrocytes by insulin. Neuropharmacology. 2014-08-23 [PMID: 25158313] (ELISA, Rat) Details: NFkB p65 ELISA (NF-kB/p65 ActivELISA kit) used for quantification of NF-kB-free p65 in the nuclear fraction of astroglial cells (Rat's astrocytoma cell line C6) which were treated or not with Streptozotocin (STZ) and/or Insulin (Fig. 5A).	ELISA	Rat
Xiao J, Xing F, Huo J et al. Lycium barbarum polysaccharides therapeutically improve hepatic functions in non-alcoholic steatohepatitis rats and cellular steatosis model Sci Rep. 2014-07-07 [PMID: 24998389] (ELISA, Rat) Details: NFkB p65 ELISA Kit / NF-kB/p65 ActivELISA kit used for quantification of the activity of transcription factor NF-kB p65 in rats from control or non-alcoholic steatohepatitis (NASH) groups treated or not with Lycium barbarum polysaccharides/LBP (Figure 5F)	ELISA	Rat
Chen SJ, Chuang LT, Chen SN. Incorporation of Eicosatrienoic Acid Exerts Mild Anti-inflammatory Properties in Murine RAW264.7 Cells Inflammation. 2014-07-04 [PMID: 24993153] (ELISA, Mouse) Details: NFkB p65 ELISA Kit / NF-kB/p65 ActivELISA-TM kit used for detection of cytosolic NF-KB p65 translocation into the nucleus in RAW264.7 cells that were incubated with 50 uM Eicosatrienoic acid (ETrA) or eicosapentaenoic acid (EPA) for 24 followed by stimulatation with LPS 2 ug/ml for 30 min (Fig 7).	ELISA	Mouse
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 2014-03-25 [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. 2014-02-18 [PMID: 24561313] (ELISA, Rat) Details: Rat brain, Fig 4.	ELISA	Rat
Umamaheswari J, Subramanian SP. Zinc-silibinin complex ameliorates oxidative stress in high fat fed low dose STZ induced type 2 diabetes in rats. Jaishanker Umamaheswari. 2015-04-01 (ELISA, Rat) Details: RelA/NFkB p65 ActivELISA kit was used for the quantification of nuclear levels of NF-kB p65 unit in pancreatic tissue homogenates of high fat fed low dose STZ induced type 2 diabetes in rats which were subjected or not to treatments with zinc-silibinin complex (Table: 5).	ELISA	Rat
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. 2014-06-06 [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
Sundaram R, Shanthi P, Sachdanandam P. Tangeretin, a polymethoxylated flavone, modulates lipid homeostasis and decreases oxidative stress by inhibiting NF-kB activation and proinflammatory cytokines in cardiac tissue of streptozotocin-induced diabetic rats. Journal of Functional Foods. 2015-06-01 (ELISA, Rat) Details: RelA/NFkB p65 ActivELISA kit was used for quantification of NF-?B free p65 in the nuclear fraction of heart tissue homogenate from streptozotocin-induced diabetic rats.	ELISA	Rat
Onodera T, Kuriyama I, Sakamoto Y et al. 5-O-Acyl plumbagins inhibit DNA polymerase activity and suppress the inflammatory response Arch. Biochem. Biophys. 2015-03-03 [PMID: 25748000] (ELISA, Mouse) Details: RelA/NFkB p65 ELISA Kit (NF-kB/p65 ActivELISA kit) was used for the measurement of NF-kB nuclear translocation in LPS/24 hour stimulated RAW 264.7 cells which were previously subjected or not to pol lambda knockdown using siRNA transfection. The signal was quantified using Molecular Devices's Vmax-K microplate reader and results were presented as relative ratio of nuclear to cytoplasmic NF-kB/p65 (Fig. 6D)	ELISA	Mouse
Abdelaziz R, Awad A, Elshaer S et al. Ameliorative effect of montelukast against STZ induced diabetic nephropathy: targeting HMGB1, TLR4, NF-kB, NLRP3 inflammasome, and autophagy pathways. Research Square 2023-04-11 (Rat)		Rat
Govindaraj J, Sorimuthu Pillai S. Rosmarinic acid modulates the antioxidant status and protects pancreatic tissues from glucolipotoxicity mediated oxidative stress in high-fat diet: streptozotocin-induced diabetic rats Mol. Cell. Biochem. 2015-06-01 [PMID: 25735949] (ELISA, Rat)	ELISA	Rat
Yang Y, Yang J, Jiang Q. The protective effect of huperzine a against hepatic ischemia reperfusion injury in mice. Transplant. Proc. 2014-06-01 [PMID: 24935330] (ELISA, Mouse) Details: Fig 4A: Hepatic tissue. Results show that NF-kB p65 activation was ncreased in ischemic mice. However, this effect could be reversed when ischemia mice were treated with huperzine A prior to the ischemia injury.	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 2014-01-01 [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. 2014-04-16 [PMID: 24737349] (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. 2014-02-21 [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. 2014-02-12 [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. 2014-01-27 [PMID: 24480038] (ELISA, Mouse) Details: Stimulated Raw264.7 cells, Fig 5A.	ELISA	Mouse
Wang SB, Jia JP. Oxymatrine attenuates diabetes-associated cognitive deficits in rats. Acta Pharmacol. Sin. 2014-03-01 [PMID: 24442148] (ELISA, Rat) Details: NF-kB activation was measured in rat cerebral cortex and hippocampus.	ELISA	Rat
Nukuzuma S, Nakamichi K, Kameoka M et al. TNF-a stimulates efficient JC virus replication in neuroblastoma cells. J. Med. Virol. 2014-01-10 [PMID: 24415534] (ELISA, Human) Details: TNF alpha stimulated and control IMR-32 neuroblastoma cells, Fig 5. The results showed that TNF alpha stimulated the activation of the NF-kB pathway.	ELISA	Human
Rastogi N, Gara RK, Trivedi R et al. (6)-Gingerol-induced myeloid leukemia cell death is initiated by reactive oxygen species and activation of miR-27b expression. Free Radic Biol Med 2013-12-27 [PMID: 24378438] (ELISA, Human)	ELISA	Human
Tsai KL, Huang PH, Kao CL et al. Aspirin attenuates Vinorelbine-induced endothelial inflammation via modulating SIRT1/AMPK axis. Biochem Pharmacol 2013-12-14 [PMID: 24345330] (ELISA, Human)	ELISA	Human
Sui X, Gao C. Huperzine A ameliorates damage induced by acute myocardial infarction in rats through antioxidant, anti-apoptotic and anti-inflammatory mechanisms. Int J Mol Med 2014-01-01 [PMID: 24190328] (ELISA)	ELISA
Zhang Y, Wang XL, Zhao J et al. Adiponectin inhibits oxidative/nitrative stress during myocardial ischemia and reperfusion via PKA signaling. Am J Physiol Endocrinol Metab 2013-12-01 [PMID: 24129398] (ELISA, Mouse)	ELISA	Mouse
Arora V, Chopra K. Possible involvement of oxido-nitrosative stress induced neuro-inflammatory cascade and monoaminergic pathway: Underpinning the correlation between nociceptive and depressive behaviour in a rodent model. J Affect Disord 2013-12-01 [PMID: 24126118] (ELISA, Rat)	ELISA	Rat
Kuhn H, Petzold K, Hammerschmidt S et al. Interaction of cyclic mechanical stretch and toll-like receptor 4-mediated innate immunity in rat alveolar type II cells. Respirology 2014-01-01 [PMID: 23796194] (ELISA, Rat)	ELISA	Rat
Lee J-J, Drakaki A, Iliopoulos D, Struhl K. MiR-27b targets PPARgamma to inhibit growth, tumor progression and the inflammatory response in neuroblastoma cells. Oncogene. 2012-08-16 [PMID: 22120719]
Oussaief L, Ramirez V, Hippocrate A et al. NF-kappaB-mediated modulation of inducible nitric oxide synthase activity controls induction of the Epstein-Barr virus productive cycle by transforming growth factor beta 1. J Virol. 2011-07-01 [PMID: 21507981]
Mhd Omar NA, Abdullah N, Kuppusamy UR et al. Nutritional Composition, Antioxidant Activities, and Antiulcer Potential of Lentinus squarrosulus (Mont.) Mycelia Extract. Evid Based Complement Alternat Med. 2011-01-01 [PMID: 21423634]
Kuhad A, Bishnoi M, Tiwari V et al. Suppression of NF-kappabeta signaling pathway by tocotrienol can prevent diabetes associated cognitive deficits. Pharmacol Biochem Behav. 2009-04-01 [PMID: 19138703] (Rat) Details: Nuclear extract from rat cerebral cortex and hippocampus (Data not shown).		Rat
Doi T, Sakoda T, Akagami T et al. Aldosterone induces interleukin-18 through endothelin-1, angiotensin II, Rho/Rho-kinase, and PPARs in cardiomyocytes. Am J Physiol Heart Circ Physiol. 2008-09-01 [PMID: 18660453] (Rat) Details: Primary cultures of rat neonatal rat cardiomyoctyes treated with various stimulators or drugs: Figs 7A-7B, Supplemental Figs. S3A-S3D.		Rat
Lin CI, Chen CN, Huang MT et al. Lysophosphatidic acid upregulates vascular endothelial growth factor-C and tube formation in human endothelial cells through LPA(1/3), COX-2, and NF-kappaB activation- and EGFR transactivation-dependent mechanisms. Cell Signal. 2008-10-01 [PMID: 18627789] (Human) Details: Human HUVEC cell culture established from freshly isolated umbilical cords, Fig 6E.		Human
Huang JR, Wu CC, Hou RC et al. Bromelain inhibits lipopolysaccharide-induced cytokine production in human THP-1 monocytes via the removal of CD14. Immunol Invest. 2008-01-01 [PMID: 18569070] Details: p65 ELISA (human THP-1 cells): Table 2. Note: Cells were treated with LPS (TLR4 ligand) which induced p65 translocation an indication of TLR activation and downstream activation of NF-kB. Bromelain, an anti-inflammatory agent, inhibited NF-kB activation.
Mangano K, Dati G, Quattrocchi C et al. Preventive and curative effects of cyclophosphamide in an animal model of Guillain Barre syndrome. J Neuroimmunol. 2008-05-30 [PMID: 18452999] (Rat) Details: Rat nerve lysates.		Rat
Kajiya M, Shiba H, Fujita T et al. Brain-derived neurotrophic factor stimulates bone/cementum-related protein gene expression in cementoblasts. J Biol Chem. 2008-06-06 [PMID: 18390540] Details: Nuclear extract of human cementoblast-like (HCEM) cells, Fig 3.
Le Negrate G, Faustin B, Welsh K et al. Salmonella secreted factor L deubiquitinase of Salmonella typhimurium inhibits NF-kB, suppresses IkappaBalpha ubiquitination and modulates innate immune responses. J Immunol. 2008-04-01 [PMID: 18354230] (Human) Details: Products cited (human HEK293N cells +/- TNF and +/- SseL plasmid transfection): 1. NK-kB/p65 ActivELISA Kit (IMK-503), Fig 2B 2. Caspase-3 pAb (IMG-5699), Fig 2A 17.		Human
Abraham WM, Ahmed A, Serebriakov I et al. Whole-body periodic acceleration modifies experimental asthma in sheep. Am J Respir Crit Care Med. 2006-10-01 [PMID: 16858016] (Sheep) Details: Nuclear extracts from sheep bronchoalveolar lung cells, Fig 7.		Sheep
Marfella R, D'Amico M, Di Filippo C et al. Increased activity of the ubiquitin-proteasome system in patients with symptomatic carotid disease is associated with enhanced inflammation and may destabilize the atherosclerotic plaque: effects of rosiglitazone treatment. J Am Coll Cardiol. 2006-06-20 [PMID: 16781372]
Leiba M, Cahalon L, Shimoni A et al. Halofuginone inhibits NF-kappaB and p38 MAPK in activated T cells. J Leukoc Biol. 2006-08-01 [PMID: 16769768] Details: Nuclear extracts from CD3 activated T cells treated with halofuginone (plant alkaloid).
Delgado M, Gonzalez-Rey E, Ganea D. The neuropeptide vasoactive intestinal peptide generates tolerogenic dendritic cells. J Immunol. 2005-12-01 [PMID: 16301637] (Human) Details: Nuclear and cytoplasmic extracts from bone marrow derived human DC cells, Fig 4.		Human
Ohsugi T, Horie R, Kumasaka T et al. In vivo antitumor activity of the NF-kappaB inhibitor dehydroxymethylepoxyquinomicin in a mouse model of adult T-cell leukemia. Carcinogenesis. 2005-08-01 [PMID: 15831528] Details: Fig 3a: nuclear extracts from HTLV-1 infected human T-cell lines MT-2 and HUT-102 cells that were treated with or without DHMEQ; (dehydroxymethylepoxyquinomicin), Fig 3b: nuclear and cytoplasmic extracts from MT-2, HUT-102 and MOLT-4 cells that were treated with or without DHMEQ. The results in Fig 3a and b showed that DHMEQ inhibited p65 nuclear translocation.
Nakagawa S, Kushiya K, Taneike I et al. Specific inhibitory action of anisodamine against a staphylococcal superantigenic toxin, toxic shock syndrome toxin 1 (TSST-1), leading to down-regulation of cytokine production and blocking of TSST-1 toxicity in mice. Clin Diagn Lab Immunol. 2005-03-01 [PMID: 15753253] Details: Nuclear lysate from THP-1 cells was stimulated with TSST-1 (toxic shock syndrome toxin 1) in the presence or the absence of anisodamine, Fig 6.
Bar-Shai M, Carmeli E, Coleman R et al. The effect of hindlimb immobilization on acid phosphatase, metalloproteinases and nuclear factor-kappaB in muscles of young and old rats. Mech Ageing Dev. 2005-02-01 [PMID: 15621209] (Rat) Details: Lysate from immobilized quadricep muscle derived from mature Female wistar rats, Fig 6.		Rat
Hecht I, Hershkoviz R, Shivtiel S et al. Heparin-disaccharide affects T cells: inhibition of NF-kappaB activation, cell migration, and modulation of intracellular signaling. J Leukoc Biol. 2004-06-01 [PMID: 15020655] Details: Nuclear and cytoplasmic extracts from CD3 activated T cells, Fig 1.
Litjens NH, Rademaker M, Ravensbergen B et al. Monomethylfumarate affects polarization of monocyte-derived dendritic cells resulting in down-regulated Th1 lymphocyte responses. Eur J Immunol. 2004-02-01 [PMID: 14768062] (Human) Details: Nuclear and cytoplasmic extracts from human dendritic cells stimulated with LPS and MMF (monomethylfumarate), Fig 4.		Human
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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.
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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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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Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric)

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