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Mouse NFκB-p65(Nuclear Factor Kappa B p65) ELISA Kit
ELK8449
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48 T $320.00
96 T $458.00
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Overview

Product name: Mouse NFκB-p65(Nuclear Factor Kappa B p65) ELISA Kit
Reactivity: Mouse
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: 11.2 pg/mL
Standard: 2000 pg/mL
Detection Range: 31.25-2000 pg/mL
Sample Type: Tissue homogenates, cell lysates and other biological fluids
Assay Length: 3.5h
Research Area: Apoptosis;
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 1.961 1.872
1000.00 1.621 1.532
500.00 1.147 1.058
250.00 0.871 0.782
125.00 0.616 0.527
62.50 0.352 0.263
31.25 0.247 0.158
0.00 0.089 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) 92-105% 98%
EDTA plasma(n=5) 83-95% 89%
Heparin plasma(n=5) 87-99% 93%

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) 92-105% 92-101% 80-94% 86-101%
EDTA plasma(n=5) 80-97% 81-92% 87-103% 82-93%
Heparin plasma(n=5) 95-104% 82-96% 92-105% 83-92%