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Mouse DEFa5(Defensin Alpha 5, Paneth Cell Specific) ELISA Kit
ELK2462
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48 T $320.00
96 T $458.00
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Overview

Product name: Mouse DEFa5(Defensin Alpha 5, Paneth Cell Specific) ELISA Kit
Reactivity: Mouse
Alternative Names: DEF-A5; DEF5; HD-5
Assay Type: Sandwich
Sensitivity: 13.1 pg/mL
Standard: 2000 pg/mL
Detection Range: 31.25-2000 pg/mL
Sample Type: serum, plasma, tissue homogenates and other biological fluids
Assay Length: 3.5h
Research Area: Infection immunity;Hepatology;Gastroenterology;
Uniprot ID: P28312
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 DEFa5. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Mouse DEFa5. 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 DEFa5, 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 DEFa5 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 2.153 2.045
1000.00 1.628 1.520
500.00 1.253 1.145
250.00 0.819 0.711
125.00 0.532 0.424
62.50 0.325 0.217
31.25 0.216 0.108
0.00 0.108 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 DEFa5 and the recovery rates were calculated by comparing the measured value to the expected amount of DEFa5 in samples.
Matrix Recovery range Average
serum(n=5) 87-95% 91%
EDTA plasma(n=5) 81-95% 88%
Heparin plasma(n=5) 83-97% 90%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of DEFa5 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) 86-100% 93-108% 80-101% 87-96%
EDTA plasma(n=5) 96-105% 95-103% 95-103% 87-98%
Heparin plasma(n=5) 85-92% 79-96% 89-101% 85-97%