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Rabbit NFκB-p65(Nuclear Factor Kappa B p65) ELISA Kit
ELK0584
Size： Price：
48 T $320.00
96 T $458.00
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Overview

Product name: Rabbit NFκB-p65(Nuclear Factor Kappa B p65) ELISA Kit
Reactivity: Rabbit
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: 18.1 pg/mL
Standard: 3000 pg/mL
Detection Range: 46.88-3000 pg/mL
Sample Type: tissue homogenates, cell lysates and other biological fluids
Assay Length: 3.5h
Research Area: Apoptosis;
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 Rabbit NFκB-p65. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rabbit 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 Rabbit 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 Rabbit 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
3000.00 2.195 2.113
1500.00 1.694 1.612
750.00 1.197 1.115
375.00 0.883 0.801
187.50 0.529 0.447
93.75 0.353 0.271
46.88 0.226 0.144
0.00 0.082 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) 83-97% 90%
EDTA plasma(n=5) 78-92% 85%
Heparin plasma(n=5) 80-95% 87%

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) 92-103% 81-92% 78-87% 91-99%
EDTA plasma(n=5) 86-98% 91-103% 85-97% 87-96%
Heparin plasma(n=5) 87-95% 85-97% 89-101% 85-97%