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
Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) Summary
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.
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
ELISA Kit (Colorimetric)
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].
Read Publications using NBP2-29661 in the following applications:
Human, mouse, rat and sheep reported in the literature.
Packaging, Storage & Formulations
Storage is content dependent.
0.02% Sodium Azide
Recombinant p65 Standard 2 vials lyophilized(0.42 ug/vial)
pNPP 4 x 5 mg
pNPP Substrate Buffer 2 x 10 ml
10X Hypotonic Lysis Buffer 10 ml
10X PBS 2 x 50 ml
100X Protease Inhibitor Cocktail (PIC) 100 ul
100 mM PMSF 10 ml
10 mM DTT (for Nuclear Lysis Buffer) 500 ul
10% Detergent Solution 10 ml
1X Nuclear Extraction Buffer 10 ml
1 M DTT (for Nuclear Extraction from tissue) 100 ul
AKP-Conjugated Secondary Ab 10 ul
Capture Antibody 200ul
Coating Buffer 2 x 10 ml
BSA 2 x 0.5 g
ELISA Plates 2
Detecting Antibody 200 ul
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)
nf kb elisa kit
nf kb p65
nfkb elisa kit
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
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
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.
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.
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).
Thangapandiyan S, Miltonprabu S, et al. Epigallocatechin gallate supplementation protects against renal injury induced by fluoride intoxication in rats: Role of Nrf2/HO-1 signaling. Toxicology Reports 2014 Mar 27 (ELISA, Rat)
Details: kidney, Fig 3D. P65 was significantly higher in fluoride-treated rats compared with control rats.
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 Jun 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.
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 Apr 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).
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 Sep 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).
FAQs for RelA/NFkB p65 ELISA Kit (NBP2-29661). (Showing 1 - 2 of 2 FAQs).
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
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.