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Mouse NFκB-p65(Nuclear Factor Kappa B p65) ELISA Kit
ELK9154
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48 T $412.00
96 T $588.00
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OverviewPublications

Product name: Mouse NFκB-p65(Nuclear Factor Kappa B p65) ELISA Kit
Reactivity: Mouse
Alternative Names: NF kappa B p65delta3; NFKB3
Assay Type: Sandwich
Sensitivity: 12.6 pg/mL
Standard: 2000 pg/mL
Detection Range: 31.25-2000 pg/mL
Sample Type: serum, plasma, tissue homogenates
Assay Length: 3.5h
Research Area: Immunology
Uniprot ID: Q04207
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 Mouse NFκB-p65. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Mouse NFκB-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 Mouse NFκB-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 Mouse NFκB-p65 in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (pg/mL) OD Corrected OD
2000.00 2.054 1.959
1000.00 1.527 1.432
500.00 1.251 1.156
250.00 0.834 0.739
125.00 0.453 0.358
62.50 0.326 0.231
31.25 0.187 0.092
0.00 0.095 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 NFκB-p65 and the recovery rates were calculated by comparing the measured value to the expected amount of NFκB-p65 in samples.
Matrix Recovery range Average
serum(n=5) 90-107% 98%
EDTA plasma(n=5) 93-107% 100%
Heparin plasma(n=5) 85-97% 91%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of NFκB-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) 86-94% 81-95% 88-97% 85-92%
EDTA plasma(n=5) 83-96% 93-102% 86-94% 90-98%
Heparin plasma(n=5) 92-106% 85-97% 96-105% 88-96%