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Mouse NFKB-p65(Nuclear Factor Kappa B p65) ELISA Kit

Citations(9)Uniprot : Q04207

Cat.No.:E-EL-M0838
Reactivity: Mouse

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96T
48T
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96T*5
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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.31-20 ng/mL
Sensitivity	0.19 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Mouse NFKB-p65 in samples. No significant cross-reactivity or interference between Mouse NFKB-p65 and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Mouse NFKB-p65 concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse NFKB-p65. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse NFKB-p65 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Mouse NFKB-p65, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Mouse NFKB-p65. You can calculate the concentration of Mouse NFKB-p65 in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse NFKB-p65 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse NFKB-p65 were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	1.05	1.94	6.71	1.15	2.11	6.27
Standard deviation	0.05	0.10	0.30	0.06	0.12	0.22
CV (%)	4.76	5.15	4.47	5.22	5.69	3.51

Recovery

The recovery of Mouse NFKB-p65 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	92-107	98
EDTA plasma (n=8)	87-101	94
Cell culture media (n=8)	88-99	94

Linearity

Samples were spiked with high concentrations of Mouse NFKB-p65 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: Q04207
Synonyms	NFKB3, p65, RelA
Research Area	Cancer, Cell Biology, Cardiovascular, Epigenetics and Nuclear Signaling, Metabolism, Microbiology, Neuroscience, Signal transduction

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

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Citations

Food & Function (2022) IF: 5.396
Black corn (Zea mays L.) whole flour improved the antioxidant capacity and prevented the adipogenesis in mice fed a high-fat diet.

DOI: 10.1039/D1FO04205J

Sample: liver
NEUROTOXICOLOGY (2021) IF: 4.294
IDO-1 inhibition protects against neuroinflammation, oxidative stress and mitochondrial dysfunction in 6-OHDA induced murine model of Parkinson’s disease

DOI: 10.1016/j.neuro.2021.03.009

Sample: Plasma,Tissue homogenate(Brain)
MOLECULES (2022) IF: 4.927
Anti-Inflammatory Effects of Auranamide and Patriscabratine—Mechanisms and In Silico Studies

DOI: 10.3390/molecules27154992

Sample: RAW 264.7 Cell
JOURNAL OF PHYSIOLOGY AND PHARMACOLOGY (2021) IF: 3.011
Rhamnetin ameliorates macrophage-mediated inflammation and pro-atherosclerosis pathways in apolipoprotein E-deficient mice

DOI: 10.26402/jpp.2021.2.10

Sample: aorta
TOXICOLOGY AND APPLIED PHARMACOLOGY (2019) IF: 3.585
Modeling of antipsychotic-induced metabolic alterations in mice: An experimental approach precluding psychosis as a predisposing factor

DOI: 10.1016/j.taap.2019.114643

PMID: 31254565
Sample: Serum
LIFE SCIENCES (2020) IF: 3.647
Berberine attenuated olanzapine-induced metabolic alterations in mice: Targeting transient receptor potential vanilloid type 1 and 3 channels

DOI: 10.1016/j.lfs.2020.117442

Sample: serum
Oncology Letters (2018) IF: 1.664
The association between inflammation, the microbiome and urethane-induced pulmonary adenocarcinoma Corrigendum in /10.3892/ol.2018.9063

DOI: 10.3892/ol.2018.8167

Sample: Serum
Molecular Medicine Reports (2018) IF: 1.922
Dioscin prevents LPS‑induced acute lung injury through inhibiting the TLR4/MyD88 signaling pathway via upregulation of HSP70

DOI: 10.3892/mmr.2018.8667

Sample: Serum,Tissue homogenate
PharmaNutrition (2022)
Digested protein from chia seed (Salvia hispanica L) prevents obesity and associated inflammation of adipose tissue in mice fed a high-fat diet

DOI: 10.1016/j.phanu.2022.100298

Sample: adipose tissue

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