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ELK Biotechnology

ELK4071

Human HBb(Hemoglobin Beta) ELISA Kit

Human HBb(Hemoglobin Beta) ELISA Kit

Size
Sensitivity
Detection range

Human HBb(Hemoglobin Beta) ELISA Kit

Overview

Product name: Human HBb(Hemoglobin Beta) ELISA Kit
Reactivity: Human
Alternative Names: HB-B; HBD; CD113t-C; Beta Globin; Spinorphin
Assay Type: Competitive Inhibition
Sensitivity: 2.94 pg/mL
Standard: 500 pg/mL
Range: 7.82-500 pg/mL
Sample Type: serum, plasma and erythrocyte lysates
Assay Length: 2h
Research Area: Hematology;
Test principle: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with Human HBb protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human HBb. 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 Human HBb 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
500.00 0.179 —
250.00 0.275 —
125.00 0.596 —
62.50 0.895 —
31.25 1.139 —
15.63 1.537 —
7.82 1.869 —
0.00 2.157 —

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 HBb and the recovery rates were calculated by comparing the measured value to the expected amount of HBb in samples.
Matrix Recovery range Average
serum(n=5) 80-92% 86%
EDTA plasma(n=5) 88-102% 95%
Heparin plasma(n=5) 90-105% 87%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of HBb 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) 92-101% 87-98% 85-94% 87-101%
EDTA plasma(n=5) 86-94% 85-95% 89-99% 85-95%
Heparin plasma(n=5) 95-102% 79-96% 87-103% 90-99%
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