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ELK Biotechnology

ELK9451

Sheep HP(Haptoglobin) ELISA Kit

Sheep HP(Haptoglobin) ELISA Kit

Sensitivity

Sheep HP(Haptoglobin) ELISA Kit

Overview

Product name: Sheep HP(Haptoglobin) ELISA Kit
Reactivity: Sheep
Alternative Names: HP; Hp2-Alpha; Alpha-2-Macroglobulin; Zonulin; Hpt; Haptoglobin
Assay Type: Sandwich
Sensitivity: 5.9 ng/mL
Standard: 1000 ng/mL
Detection Range: 15.63-1000 ng/mL
Sample Type: serum, plasma and other biological fluids
Assay Length: 3.5h
Research Area: Infection immunity;Hematology;
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 Sheep HP. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Sheep HP. 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 Sheep HP, 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 Sheep HP 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
1000.00 2.135 2.042
500.00 1.599 1.506
250.00 1.120 1.027
125.00 0.897 0.804
62.50 0.518 0.425
31.25 0.335 0.242
15.63 0.226 0.133
0.00 0.093 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 HP and the recovery rates were calculated by comparing the measured value to the expected amount of HP in samples.
Matrix Recovery range Average
serum(n=5) 95-107% 101%
EDTA plasma(n=5) 80-97% 88%
Heparin plasma(n=5) 82-94% 88%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of HP 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) 81-98% 78-90% 91-99% 97-103%
EDTA plasma(n=5) 82-94% 86-96% 81-97% 84-97%
Heparin plasma(n=5) 88-97% 85-92% 79-96% 87-103%
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