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Bio Reagents & Consumables
ELISA (Enzyme-linked Immunosorbent Assay)
ELK1438 Human HMGB-1 (HighMobility Group Protein B1) ELISA Kit
Human ELISA Kit
Brand: ELK Biotechnology
Available
Free delivery within Singapore only
Overview
| Product name: | Human HMGB-1(HighMobility 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: | 28.3 pg/mL |
| Standard: | 4000 pg/mL |
| Range: | 62.5-4000 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 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 (ng/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% |
Specification :
| Details | Kindly refer to the "Overview" tab. |
