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Human NFκB-p65(Nuclear Factor Kappa B p65) Microsample ELISA Kit
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ELK1692MS
Size： Price：
48 T $320.00
96 T $458.00
96 T*5 $1947.00
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Overview

Product name: Human NFκB-p65(Nuclear Factor Kappa B p65) Microsample 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: 0.053 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 1.905 1.817
5.00 1.568 1.480
2.50 1.172 1.084
1.25 0.854 0.766
0.63 0.549 0.461
0.31 0.372 0.284
0.16 0.184 0.096
0.00 0.088 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) 80-111% 96%
EDTA plasma(n=5) 87-99% 93%
Heparin plasma(n=5) 94-116% 105%

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) 93-102% 94-119% 87-108% 82-101%
EDTA plasma(n=5) 83-116% 83-111% 85-97% 82-92%
Heparin plasma(n=5) 86-99% 94-107% 92-111% 81-96%

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