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ELK Biotechnology

ELK9475

Horse Irisin ELISA Kit

Horse Irisin ELISA Kit

Sensitivity

Horse Irisin ELISA Kit

Overview

Product name: Horse Irisin ELISA Kit
Reactivity: Horse
Alternative Names: FRCP2; Irisin; Fibronectin type III repeat-containing protein 2; FNDC5; Fibronectin Type III Domain Containing Protein 5
Assay Type: Sandwich
Sensitivity: 28.4 pg/mL
Standard: 4000 pg/mL
Detection Range: 62.5-4000 pg/mL
Sample Type: serum, plasma, tissue homogenates and other biological fluids
Assay Length: 3.5h
Research Area: CD & Adhesion molecule;
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 Horse Irisin. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Horse Irisin. 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 Horse Irisin, 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 Horse Irisin 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
4000.00 2.266 2.172
2000.00 1.612 1.518
1000.00 1.282 1.188
500.00 0.892 0.798
250.00 0.513 0.419
125.00 0.355 0.261
62.50 0.236 0.142
0.00 0.094 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 Irisin and the recovery rates were calculated by comparing the measured value to the expected amount of Irisin in samples.
Matrix Recovery range Average
serum(n=5) 81-97% 89%
EDTA plasma(n=5) 94-107% 100%
Heparin plasma(n=5) 83-95% 89%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Irisin 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) 93-103% 94-106% 91-101% 95-103%
EDTA plasma(n=5) 88-102% 84-97% 89-107% 85-92%
Heparin plasma(n=5) 90-96% 92-103% 93-105% 88-101%
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