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Dog HMGB-1(High Mobility Group Protein B1) ELISA Kit
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ELK10709
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48 T $412.00
96 T $588.00
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Overview

Product name: Dog HMGB-1(High Mobility 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: 0.064 ng/mL
Standard: 10 ng/mL
Detection Range: 0.16-10 ng/mL
Sample type: serum, plasma and other biological fluids
Assay length: 3.5h
Research Area: Infection immunity;
Uniprot ID: Q6YKA4
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
10.00 2.154 2.059
5.00 1.576 1.481
2.50 1.114 1.019
1.25 0.847 0.752
0.63 0.518 0.423
0.32 0.352 0.257
0.16 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%