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ELK Biotechnology

ELK9584

General F-TESTO(FreeTestosterone) ELISA Kit

General F-TESTO(FreeTestosterone) ELISA Kit

Sensitivity

General F-TESTO(FreeTestosterone) ELISA Kit

Overview

Product name: F-TESTO(FreeTestosterone) ELISA Kit
Reactivity: General
Alternative Names: F-TESTO
Assay Type: Competitive Inhibition
Sensitivity: 18.75 pg/mL
Standard: 2000 pg/mL
Detection Range: 31.25-2000 pg/mL
Sample Type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 2h
Research Area: Endocrinology;Reproductive science;Hormone metabolism;
Test principle: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with F-TESTO protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to F-TESTO. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. 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 F-TESTO 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
2000.00 0.198 —
1000.00 0.385 —
500.00 0.621 —
250.00 0.842 —
125.00 1.275 —
62.50 1.565 —
31.25 1.787 —
0.00 2.057 —

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 F-TESTO and the recovery rates were calculated by comparing the measured value to the expected amount of F-TESTO in samples.
Matrix Recovery range Average
serum(n=5) 81-95% 88%
EDTA plasma(n=5) 83-95% 89%
Heparin plasma(n=5) 89-103% 96%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of F-TESTO 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) 93-104% 87-103% 91-102% 87-102%
EDTA plasma(n=5) 87-96% 92-101% 85-98% 97-104%
Heparin plasma(n=5) 92-101% 82-96% 95-102% 95-104%
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