Mouse nuclear factor-κB p65,NF-κB p65 ELISA Kit

Code CSB-E08789m
Size 96T,5×96T,10×96T How to order?
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Product Details

Description

The mouse NF-κB subunit p65 ELISA kit is suitable for the quantitative determination of mouse p65 in different sample types, including serum, plasma, and tissue homogenates. This assay employs the bi-antibody sandwich technique and enzyme-substrate chromogenic reaction to quantify antigen levels in the sample. The amount of synthesized colored products is positively related to the analyte of interest in the sample.

p65, also called RELA, is a member of the NF-κB family of transcriptional regulatory proteins that works as the activating component of the p65/p50 heterodimer. It plays an essential role in cell survival as p65-deficient mice do not survive. Nuclear translocation of the p65/p50 heterodimer is essential for NF-κB signaling. On activation of TNFR1, the TRAF2/5 activates the IKK complex, resulting in IκBα phosphorylation and subsequent ubiquitin-dependent proteasomal degradation. This relieves the NF-κB p65/p50 heterodimer from cytosolic retention, leading to nuclear translocation and target gene activation. Acetylation of p65 affects its biological function. Acetylation on lysine residue 221 is crucial for DNA binding and for inhibiting the interaction between p65 and IκBα.

Target Name v-rel reticuloendotheliosis viral oncogene homolog A (avian)
Alternative Names Rela ELISA Kit; Nfkb3 ELISA Kit; Transcription factor p65 ELISA Kit; Nuclear factor NF-kappa-B p65 subunit ELISA Kit; Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 ELISA Kit
Abbreviation RELA
Uniprot No. Q04207
Species Mus musculus (Mouse)
Sample Types serum, plasma, tissue homogenates
Detection Range 31.25 pg/mL-2000 pg/mL
Sensitivity 7.8 pg/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Immunology
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%        
Three samples of known concentration were tested twenty times on one plate to assess.    
Inter-assay Precision (Precision between assays): CV%<10%        
Three samples of known concentration were tested in twenty assays to assess.      
               
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of mouse NF-κB p65 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.  
  Sample Serum(n=4)    
1:1 Average % 96    
Range % 92-100    
1:2 Average % 84    
Range % 80-88    
1:4 Average % 93    
Range % 89-97    
1:8 Average % 89    
Range % 84-93    
Recovery
The recovery of mouse NF-κB p65 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.  
 
Sample Type Average % Recovery Range    
Serum (n=5) 104 100-110    
EDTA plasma (n=4) 95 90-100    
               
               
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.  
 
pg/ml OD1 OD2 Average Corrected    
2000 2.659 2.600 2.630 2.451    
1000 2.042 1.935 1.989 1.810    
500 1.211 1.270 1.241 1.062    
250 0.736 0.724 0.730 0.551    
125 0.394 0.408 0.401 0.222    
62.5 0.307 0.311 0.309 0.130    
31.25 0.258 0.247 0.253 0.074    
0 0.175 0.182 0.179      
ELISA Data Analysis Watch ELISA data processing video & download Curve Expert if needed
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

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Target Background

Function
(From Uniprot)
NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52. The heterodimeric RELA-NFKB1 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. The NF-kappa-B heterodimeric RELA-NFKB1 and RELA-REL complexes, for instance, function as transcriptional activators. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. The inhibitory effect of I-kappa-B on NF-kappa-B through retention in the cytoplasm is exerted primarily through the interaction with RELA. RELA shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Beside its activity as a direct transcriptional activator, it is also able to modulate promoters accessibility to transcription factors and thereby indirectly regulate gene expression. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1. Essential for cytokine gene expression in T-cells. The NF-kappa-B homodimeric RELA-RELA complex appears to be involved in invasin-mediated activation of IL-8 expression.
Gene References into Functions
  1. This study reports here an X-ray crystal structure of homodimers comprising the RelA DNA-binding domain containing the Rel homology region (RHR) in NF-kappa B bound to an E-selectin promoter fragment with tandem kappa B sites. PMID: 28935669
  2. p65/NF-kappaB controls ankyrin-G levels in axon initial segment via a negative feedback loop. PMID: 28181483
  3. During skeletal development, homozygous knockout of transcription factor RelA (Rela) leads to impaired growth through ectopic apoptosis of chondrocytes, whereas heterozygous knockout of Rela does not alter growth. PMID: 27830706
  4. NF-kappa B p65 transcriptional activity is regulated by platelet-activating factor in macrophages. PMID: 27554194
  5. Thalidomide potentiates etoposide-induced apoptosis in murine neuroblastoma by suppressing NF-kappaB. PMID: 29423589
  6. Pudilan xiaoyan oral liquid prevents LPS-induced respiratory in fl ammation via effects on TLR4/NF-kappaB signaling. PMID: 28689797
  7. Catalase ameliorates diabetes-induced cardiomyopathy through reduced RelA-mediated transcription of BECN1. PMID: 28643395
  8. SOD2 expression is ATM- and RelA-dependent, ATM knockdown renders cells sensitive to pro-oxidant exposure, and SOD mimetics partially rescue this sensitivity. Mice with germline deletion of Atm fail to develop mature mammary glands, but using a conditional knockout approach, we determined that Atm deletion significantly diminished the expression of Sod2. PMID: 28849346
  9. RelA-BRD4 signaling in nonciliated bronchiolar epithelial cells mediates neutrophilic airway inflammation and Respiratory Syncytial disease severity. PMID: 29593031
  10. A significant decline of p65 level was observed in the brain tissues of hamsters infected with scrapie agent 263K at terminal stage. p65 colocalized with neuronal nuclear protein positive cells but not with glial fibrillary acidic protein positive cells. PMID: 28783945
  11. these findings suggest that BRD7 has an anti-inflammatory role during early acute inflammation by inhibiting activation of the NF-small ka, CyrillicB signaling pathway, which provides evidence to aid in understanding the therapeutic effects of BRD7 on inflammatory diseases PMID: 27374794
  12. NF-kappaB1 modifies acute inflammatory renal injury but does not influence chronic fibrotic injury PMID: 28617440
  13. beta-catenin and p65 are activated in separate cellular compartments during liver regeneration, with p65 activity in nonparenchymal compartment contributing to the activation of hepatocyte beta-catenin, cyclin D1 expression, and subsequent proliferation PMID: 28474571
  14. the enhancement of stat3 phosphorylation and decrease of LPS-induced p65 translocation were achieved by nicotine treatment..this study revealed that TIPE2 upregulation and stat3 phosphorylation contribute to nicotine-mediated anti-inflammation effect, indicating that TIPE2 and stat3 might be potential molecules for dealing with inflammation-associated diseases. PMID: 28766689
  15. The data demonstrate that miR-125b regulates nasopharyngeal carcinoma cell proliferation and apoptosis by targeting A20/NF-kappaB signaling pathway, and miR-125b acts as oncogene, whereas A20 functions as tumor suppressor. PMID: 28569771
  16. TLR4-NF-kappaB signal pathway is inhibited by paeonol in endotoxin-induced acute kidney injury PMID: 27027358
  17. Type III secretion system of enteropathogenic Escherichia coli is necessary and sufficient for host NF-kappaB activation. PMID: 28671993
  18. TNF-alpha and the transcription factor nuclear factor kappa B (NF-kappaB) p65 subunit were both upregulated in bleomycin-induced fibrotic lung tissue. PMID: 28731277
  19. These findings suggested that USP14 induces NF-kappaB activity and ERK1/2 phosphorylation triggered by microbial infection. PMID: 28364380
  20. this study demonstrates that phosphorylation of p65 serine 467 augment NF-kappaB activity and exacerbates various deleterious effects of overnutrition in mice PMID: 28723419
  21. RORalpha/gamma are important therapeutic targets for cutaneous inflammation; RORa and RORg have roles in inflammation in mouse models of atopic dermatitis and acute irritant dermititis PMID: 28774591
  22. Expansion of myeloid-derived suppressor cells with aging in the bone marrow of mice through a Rela-dependent mechanism. PMID: 28229533
  23. p65 expression correlates with elevated total cellular levels of STAT3 and STAT1 and supports activation of these transcription factors. PMID: 28089769
  24. Acanthopanax senticosus polysaccharide pretreatment may be associated with inhibition of the NF-kappaB/MLCK pathway and concomitant amelioration of LPS-induced tight junction dysfunction of intestinal epithelium in endotoxemia PMID: 28405145
  25. Data suggest that activation of NFkappaB/RelA can be moderated by dietary factors; here, naringenin (an anti-inflammatory, antioxidant flavanone dietary supplement) reduces lipopolysaccharide-induced inflammatory pain, mechanical hyperalgesia, and thermal hyperalgesia; naringenin reduces mononuclear-leukocyte/neutrophil/macrophage recruitment by inhibiting NFkappaB/RelA activation. PMID: 27260463
  26. results suggest that Ser(534) phosphorylation of p65 in mice (and, by extension, Ser(536) phosphorylation of human p65) is not required for its nuclear translocation, but instead inhibits NF-kappaB signaling to prevent deleterious inflammation. PMID: 27555662
  27. Angiotensin II regulates dendritic cells through activation of p65 NF-kappaB, ERK1, ERK2 and STAT1 pathways. PMID: 28723692
  28. RelA has a role in regulating OIS in preneoplastic lesions; the RelA/CXCL1/CXCR2 axis is an essential mechanism of tumor surveillance in pancreatic ductal adenocarcinoma PMID: 27454298
  29. This study suggests that mTOR activity in hepatocytes decreases hepatic vulnerability to injury through a mechanism dependent on NF-kappaB proinflammatory cytokine signaling pathway in both normal and steatotic liver. PMID: 28356345
  30. this study demonstrates that lipopolysaccharides, TNF-alpha, and viral infection, all of which induce robust inflammatory responses in naturally differentiated cells, failed to activate NF-kappaB, the key transcription factor that mediates inflammatory responses, and were unable to induce the expression of inflammatory genes in mouse embryonic stem cells. Similar results were obtained in human embryonic PMID: 28130495
  31. this study illustrates a significant role of RelA in mediating the corneal neovascularization PMID: 27062711
  32. Consistent with these effects, RelA T505A mice exhibit earlier onset of cancer in the N-nitrosodiethylamine model of hepatocellular carcinoma. These data reveal a critical pathway controlling NF-kappaB function in the liver that acts to suppress the tumour-promoting activities of RelA. PMID: 26853469
  33. blockage of NF-kappaB p65 and/or MAPK p38 with their specific inhibitors strongly attenuated B7-H3-amplified inflammatory response with significantly reduced proinflammatory cytokine and chemokine production, and markedly ameliorated B7-H3-exacerbated disruption of blood-brain barrier and severity of disease status in S. pneumoniae-infected mice. PMID: 28141831
  34. our data show that Substance P (SP) treatment might be associated with anti-inflammatory effects in LPS-stimulated RAW 264.7 cells by suppressing NF-kappaB (p65)activation and inducing HO-1 expression. PMID: 27907187
  35. this study shows that LPS-induced pro-inflammatory cytokine production is ameliorated in p65-deficient bone marrow-derived macrophages; however, p65-deficient 'activated' peritoneal macrophages exhibited elevated IL-1beta and IL-6 PMID: 27932520
  36. tis study reveals the role of NF-kB signaling pathway in otitis media pathogenesis, and more specifically in mucosal proliferation PMID: 27655045
  37. Cardiac myocyte subpopulations differ in their NF-kappaB subcellular localization and identify the cis-Golgi as a cardiac myocyte-specific host compartment. PMID: 28043025
  38. TCF-4 as a co-activator of p65 in the potentiation of proinflammatory cytokine production in macrophages and aggravation of high-fat diet induced chronic inflammation and insulin resistance in mice. PMID: 27129186
  39. elevation of O-GlcNAcylation by overexpression of OGT increased the expression of p300, IKKalpha, and IKKbeta and promoted IKK-mediated activating phosphorylation of p65 at Ser-536, contributing to NF-kappaB activation. In addition, we also identified phosphorylation of p65 at Thr-308, which might impair the O-GlcNAcylation of p65 at Thr-305. PMID: 28416608
  40. this study shows that curcumin reduces the progression of progression of non-alcoholic steatohepatitis and liver damage, which may act via inhibiting HMGB1-NF-kappaB translocation PMID: 28110063
  41. long term Cryptococcus neoformansinfection induces stable nuclear localization of p65 and IkappaBalpha proteins in the absence of additional pro-inflammatory stimuli. In this case, nuclear localization of p65 is not accompanied by TNFalpha or inducible NOS (iNOS) expression. PMID: 27231343
  42. this study shows that genipin protects against acute lung injury through inhibiting NF-kappaB signaling pathway PMID: 27262946
  43. this study shows that hyperin protects against cisplatin-induced acute kidney injury by inhibiting NF-KB signaling PMID: 27764742
  44. this paper shows that isocyperol mediates its anti-inflammatory activity via suppression of the NF-kB pathway PMID: 27240136
  45. The results indicate that MHE exerts anti-inflammatory effects by suppressing inflammatory mediator production via NF-kappaB and MAPK signaling pathways inhibition and induction of HO-1 expression in macrophages. Therefore, our results suggest the potential value of MHE as an inflammatory therapeutic agent developed from a natural substance. PMID: 27338335
  46. This study shows that Astragaloside IV protects against the progression of renal fibrosis by inhibiting inflammation via the TLR4/NF-small ka, CyrillicB signaling pathway PMID: 27855303
  47. this study shows that attenuating of atherosclerosis by berberine and 8-cetylberberine attributes to inhibition of the translocation of NF-kappaB to the nucleus PMID: 28024280
  48. These findings showing that RelA controls Treg stability and promotes the competitive fitness of effector Tregs highlight the importance of RelA activity in peripheral Treg induced tolerance. PMID: 27068879
  49. this paper shows that inhibition of autophagic flux regulates the alternation of microglial phenotype in cerebral ischemia by modulating the balance between NF-kappaB and CREB PMID: 27474951
  50. Data indicate that heme oxygenase-1 (HO-1)-mediated downregulation of NF-kappa B p65 was involved in the anti-psoriatic effect of Terminalia chebulanin (TC). PMID: 27383847

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Subcellular Location Nucleus. Cytoplasm.
Database Links

KEGG: mmu:19697

STRING: 10090.ENSMUSP00000025867

UniGene: Mm.249966

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