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ELK Biotechnology

ELK9589

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

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

Sensitivity

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

Overview

Product name: Human NFKB-p65(Nuclear Factor Kappa B p65) ELISA Kit
Reactivity: Human
Alternative Names: NFKB3; p65; RelA
Assay Type: Sandwich
Sensitivity: 0.10 ng/mL
Standard: 10 ng/mL
Detection Range: 0.16-10 ng/mL
Sample Type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 3.5h
Research Area: Signal transduction;
Test principle: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Human NFKB-p65. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human NFKB-p65. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Human NFKB-p65, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Human NFKB-p65 in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (ng/mL) OD Corrected OD
10.00 1.956 1.858
5.00 1.674 1.576
2.50 1.174 1.076
1.25 0.797 0.699
0.63 0.555 0.457
0.32 0.377 0.279
0.16 0.236 0.138
0.00 0.098 0.000

Precision

Intra-assay Precision (Precision within an assay)：CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays)：CV%<10%

Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.

Recovery

Matrices listed below were spiked with certain level of recombinant NFKB-p65 and the recovery rates were calculated by comparing the measured value to the expected amount of NFKB-p65 in samples.
Matrix Recovery range Average
serum(n=5) 96-107% 101%
EDTA plasma(n=5) 79-93% 86%
Heparin plasma(n=5) 80-97% 88%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of NFKB-p65 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Matrix 1:2 1:4 1:8 1:16
serum(n=5) 89-98% 95-103% 99-106% 78-92%
EDTA plasma(n=5) 82-96% 87-103% 91-97% 81-93%
Heparin plasma(n=5) 78-91% 87-98% 92-101% 96-105%
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