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ELK Biotechnology

ELK9766

Dog HA(Histamine) ELISA Kit

Dog HA(Histamine) ELISA Kit

Sensitivity

Dog HA(Histamine) ELISA Kit

Overview

Product name: Dog HA(Histamine) ELISA Kit
Reactivity: Dog
Alternative Names: Histamin
Assay Type: Competitive Inhibition
Sensitivity: 0.51 ng/mL
Standard: 100 ng/mL
Detection Range: 1.57-100 ng/mL
Sample Type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 2h
Research Area: Signal transduction;Infection immunity;
Test principle: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with Dog HA protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Dog HA. 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 Dog HA 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
100.00 0.199 —
50.00 0.397 —
25.00 0.611 —
12.50 0.897 —
6.25 1.297 —
3.13 1.578 —
1.57 1.869 —
0.00 2.112 —

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 HA and the recovery rates were calculated by comparing the measured value to the expected amount of HA in samples.
Matrix Recovery range Average
serum(n=5) 85-99% 92%
EDTA plasma(n=5) 90-105% 97%
Heparin plasma(n=5) 78-96% 87%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of HA 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) 82-95% 89-102% 85-92% 96-104%
EDTA plasma(n=5) 97-106% 86-97% 92-99% 79-96%
Heparin plasma(n=5) 89-101% 88-97% 97-105% 81-93%
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