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Human NFκB-p65(Nuclear Factor Kappa B p65) ELISA Kit
ELK1692
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48 T $320.00
96 T $458.00
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OverviewPublications

Product name: Human NFκB-p65(Nuclear Factor Kappa B p65) ELISA Kit
Reactivity: Human
Alternative Names: NFKB3; p65; REL-A; Nuclear Factor Of Kappa Light Polypeptide Gene Enhancer In B-Cells 3; Transcription Factor P65; Nuclear factor NF-kappa-B p65 subunit; RELA; V-Rel Reticuloendotheliosis Viral Oncogene Homolog A
Assay Type: Sandwich
Sensitivity: 0.055 ng/mL
Standard: 10 ng/mL
Detection Range: 0.16-10 ng/mL
Sample Type: tissue homogenates, cell lysates and other biological fluids
Assay Length: 3.5h
Research Area: Apoptosis;
Uniprot ID: Q04206
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 NFκB-p65. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human 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 Human 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 Human NFκB-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 1.961 1.875
5.00 1.587 1.501
2.50 1.185 1.099
1.25 0.869 0.783
0.63 0.549 0.463
0.32 0.366 0.280
0.16 0.179 0.093
0.00 0.086 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) 80-92% 86%
EDTA plasma(n=5) 83-97% 90%
Heparin plasma(n=5) 86-99% 92%

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) 86-93% 93-101% 85-98% 90-101%
EDTA plasma(n=5) 89-99% 83-99% 98-105% 96-101%
Heparin plasma(n=5) 85-92% 79-96% 89-101% 85-97%