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ELK Biotechnology

ELK9768

Dog HbA1c(Glycated Hemoglobin A1c) ELISA Kit

Dog HbA1c(Glycated Hemoglobin A1c) ELISA Kit

Sensitivity

Dog HbA1c(Glycated Hemoglobin A1c) ELISA Kit

Overview

Product name: Dog HbA1c(Glycated Hemoglobin A1c) ELISA Kit
Reactivity: Dog
Alternative Names: Glycosylated Hemoglobin; Hemoglobin A1c; Hb1c; HbAIc; HbAIc
Assay Type: Competitive Inhibition
Sensitivity: 4.25 μg/mL
Standard: 1000 μg/mL
Detection Range: 15.63-1000 μg/mL
Sample Type: plasma and erythrocyte lysates
Assay Length: 2h
Research Area: Diagnosis of diabetes mellitus
Test principle: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with Dog HbA1c protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Dog HbA1c. 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 HbA1c 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 0.185 —
500.00 0.336 —
250.00 0.557 —
125.00 0.899 —
62.50 1.236 —
31.25 1.537 —
15.63 1.733 —
0.00 2.213 —

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 HbA1c and the recovery rates were calculated by comparing the measured value to the expected amount of HbA1c in samples.
Matrix Recovery range Average
serum(n=5) 90-105% 97%
EDTA plasma(n=5) 88-102% 95%
Heparin plasma(n=5) 94-107% 100%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of HbA1c 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) 82-90% 92-101% 85-92% 87-103%
EDTA plasma(n=5) 95-103% 97-105% 88-97% 97-105%
Heparin plasma(n=5) 87-94% 93-101% 88-95% 96-101%
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