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ELK Biotechnology

ELK5992

Dog HMG1(High Mobility Group Protein 1) ELISA Kit

Dog HMG1(High Mobility Group Protein 1) ELISA Kit

Size
Sensitivity
Detection range

Dog HMG1(High Mobility Group Protein 1) ELISA Kit

Overview

Product name: Dog HMGB-1(HighMobility Group Protein B1) ELISA Kit
Reactivity: Dog
Alternative Names: HMGB1; HMG3; SBP1; Sulfoglucuronyl Carbohydrate Binding Protein; Amphoterin; High Mobility Group Box 1 Protein; HMG1; High Mobility Group Protein 1
Assay Type: Sandwich
Sensitivity: 2.73 pg/mL
Standard: 400 pg/mL
Detection Range: 6.25-400 pg/mL
Sample Type: serum, plasma and other biological fluids
Assay Length: 3.5h
Research Area: Infection immunity;
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 Dog HMGB-1. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Dog HMGB-1. 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 Dog HMGB-1, 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 Dog HMGB-1 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
400.00 2.154 2.059
200.00 1.576 1.481
100.00 1.114 1.019
50.00 0.847 0.752
25.00 0.518 0.423
12.50 0.352 0.257
6.25 0.225 0.130
0.00 0.095 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 HMGB-1 and the recovery rates were calculated by comparing the measured value to the expected amount of HMGB-1 in samples.
Matrix Recovery range Average
serum(n=5) 82-97% 89%
EDTA plasma(n=5) 84-93% 87%
Heparin plasma(n=5) 95-108% 101%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of HMGB-1 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) 79-94% 87-96% 78-93% 82-90%
EDTA plasma(n=5) 98-107% 90-99% 88-97% 85-97%
Heparin plasma(n=5) 92-103% 93-105% 88-101% 97-106%
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