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Human HMGB-1(High Mobility Group Protein B1) ELISA Kit
ELK1438
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48 T $320.00
96 T $458.00
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Product name: Human HMGB-1(High Mobility Group Protein B1) ELISA Kit
Reactivity: Human
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: 26.9 pg/mL
Standard: 4000 pg/mL
Detection Range: 62.5-4000 pg/mL
Sample Type: serum, plasma and other biological fluids
Assay Length: 3.5h
Research Area: Infection immunity;
Uniprot ID: P09429
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 Human HMGB-1. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human 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 Human 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 Human HMGB-1 in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (pg/mL) OD Corrected OD
4000.00 1.961 1.868
2000.00 1.436 1.343
1000.00 1.196 1.103
500.00 0.841 0.748
250.00 0.614 0.521
125.00 0.415 0.322
62.50 0.225 0.132
0.00 0.093 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) 80-97% 88%
EDTA plasma(n=5) 85-101% 93%
Heparin plasma(n=5) 83-97% 90%

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) 91-99% 83-89% 88-104% 97-103%
EDTA plasma(n=5) 85-104% 83-96% 95-103% 88-95%
Heparin plasma(n=5) 86-93% 93-101% 86-97% 90-101%