| Product Name |
Rat NFKB-p65(NuclearFactor Kappa B p65) ELISA Kit |
| Catalogue No. |
3961ELK |
| Alternative Names: |
B-Cell CLL/Lymphoma 2 |
| Assay Type |
Sandwich |
| Sensitivity |
0.058 ng/mL |
| Range |
0.16-10 ng/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 NuclearFactor Kappa B p65(NFKB-p65). Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to NuclearFactor Kappa B p65(NFKB-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 NuclearFactor Kappa B p65(NFKB-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 NuclearFactor Kappa B p65(NFKB-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 |
1.978 |
1.891 |
| 5.00 |
1.536 |
1.449 |
| 2.50 |
1.184 |
1.097 |
| 1.25 |
0.841 |
0.754 |
| 0.63 |
0.554 |
0.467 |
| 0.32 |
0.381 |
0.294 |
| 0.16 |
0.190 |
0.103 |
| 0.00 |
0.087 |
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 NFKB-p65 and the recovery rates were calculated by comparing the measured value to the expected amount of NFKB-p65 in samples.
| Matrix |
Recovery range |
Average |
| serum(n=5) |
83-97% |
90% |
| EDTA plasma(n=5) |
81-95% |
88% |
| Heparin plasma(n=5) |
85-95% |
90% |
|
| Linearity |
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of NFKB-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% |
93-102% |
89-97% |
85-94% |
| EDTA plasma(n=5) |
91-99% |
83-97% |
87-105% |
87-99% |
| Heparin plasma(n=5) |
85-96% |
87-101% |
89-102% |
85-98% |
|
| Note |
For Research Use Only |
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