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ELK Biotechnology
SKU:ELK3025
Human C5b-9(Terminal Complement Complex C5b-9) ELISA Kit
Human C5b-9(Terminal Complement Complex C5b-9) ELISA Kit
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Human C5b-9(Terminal Complement Complex C5b-9) ELISA Kit
Overview
| Product name: | Human sC5b-9(Soluble Terminal Complement Complex C5b-9) ELISA Kit |
| Reactivity: | Human |
| Alternative Names: | MAC; Membrane Attack Complex; C5b-9; Terminal Complement Complex C5b-9; C5b9 |
| Assay Type: | Sandwich |
| Sensitivity: | 0.51 ng/mL |
| Standard: | 80 ng/mL |
| Range: | 1.25-80 ng/mL |
| Sample Type: | serum, plasma and other biological fluids |
| Assay Length: | 3.5h |
| Research Area: | Signal transduction;Metabolic pathway;Apoptosis;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 sC5b-9. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human sC5b-9. 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 sC5b-9, 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 sC5b-9 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 |
|---|---|---|
| 80.00 | 2.274 | 2.196 |
| 40.00 | 1.761 | 1.683 |
| 20.00 | 1.528 | 1.450 |
| 10.00 | 0.951 | 0.873 |
| 5.00 | 0.503 | 0.425 |
| 2.50 | 0.351 | 0.273 |
| 1.25 | 0.252 | 0.174 |
| 0.00 | 0.078 | 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 sC5b-9 and the recovery rates were calculated by comparing the measured value to the expected amount of sC5b-9 in samples.
| Matrix | Recovery range | Average |
|---|---|---|
| serum(n=5) | 81-95% | 88% |
| EDTA plasma(n=5) | 83-95% | 89% |
| Heparin plasma(n=5) | 87-97% | 90% |
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of sC5b-9 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) | 97-105% | 86-98% | 84-93% | 89-102% |
| EDTA plasma(n=5) | 87-102% | 85-94% | 87-101% | 89-99% |
| Heparin plasma(n=5) | 85-101% | 78-96% | 79-98% | 93-102% |
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