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Rat TRAF6(TNF Receptor Associated Factor 6) ELISA Kit
ELK0501
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48 T $320.00
96 T $458.00
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Overview

Product name: Rat TRAF6(TNF Receptor Associated Factor 6) ELISA Kit
Reactivity: Rat
Alternative Names: RNF85; E3 ubiquitin-protein ligase TRAF6; Interleukin-1 signal transducer
Assay Type: Sandwich
Sensitivity: 8.1 pg/mL
Standard: 1000 pg/mL
Detection Range: 15.63-1000 pg/mL
Sample Type: Tissue homogenates, cell lysates and other biological fluids.
Assay Length: 3.5h
Research Area: Tumor immunity;
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 Rat TRAF6. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat TRAF6. 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 Rat TRAF6, 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 Rat TRAF6 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
1000.00 1.948 1.862
500.00 1.612 1.526
250.00 1.285 1.199
125.00 0.912 0.826
62.50 0.627 0.541
31.25 0.315 0.229
15.63 0.158 0.072
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 TRAF6 and the recovery rates were calculated by comparing the measured value to the expected amount of TRAF6 in samples.
Matrix Recovery range Average
serum(n=5) 90-103% 96%
EDTA plasma(n=5) 85-97% 91%
Heparin plasma(n=5) 82-95% 88%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of TRAF6 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) 84-99% 81-95% 98-107% 96-101%
EDTA plasma(n=5) 89-101% 87-98% 92-104% 82-96%
Heparin plasma(n=5) 93-102% 80-93% 91-100% 88-95%