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Assay Type
Solid Phase Sandwich ELISA
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Format
96-well strip plate
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Assay Length
4.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (100 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL), Citrate Plasma (100 uL)
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Sensitivity
0.7 pg/mL
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Assay Range
3.1 - 300 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Specificity
Natural and recombinant human IL-6
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Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
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Interference
No significant interference observed with available related molecules.
Control Available
QC01-1, Quantikine Immunoassay Control Group 1 -
Product Summary
The Quantikine Human IL-6 Immunoassay is a 4.5 hour solid phase ELISA designed to measure IL-6 in cell culture supernates, serum, and plasma. It contains recombinant human IL-6 and antibodies raised against recombinant human IL-6 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural IL-6 showed linear curves that were parallel to the standard curves obtained using the E. coli-expressed Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-6.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 15.8 | 95.6 | 179 | 16.4 | 98.8 | 188 |
| Standard Deviation | 0.7 | 3 | 3.1 | 0.6 | 2.5 | 3.7 |
| CV% | 4.4 | 3.1 | 1.7 | 3.7 | 2.5 | 2 |
Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 16.8 | 97.7 | 186 | 17.2 | 101 | 191 |
| Standard Deviation | 0.7 | 1.6 | 3.8 | 1.1 | 3.3 | 7.2 |
| CV% | 4.2 | 1.6 | 2 | 6.4 | 3.3 | 3.8 |
Recovery
The recovery of human IL-6 spiked to three different levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Media (n=5) | 98 | 94-103 |
| Citrate Plasma (n=5) | 91 | 82-95 |
| EDTA Plasma (n=5) | 95 | 84-101 |
| Heparin Plasma (n=5) | 90 | 88-98 |
| Serum (n=5) | 93 | 86-99 |
Linearity
To assess the linearity of the assay, four samples were spiked with high concentrations of IL-6 in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Background: IL-6
IL-6 (Interleukin-6) is a pleiotropic cytokine that acts in the acute phase reaction, inflammation, hematopoiesis, bone metabolism, and cancer progression. It contributes to chronic inflammation in obesity, insulin resistance, inflammatory bowel disease, arthritis, sepsis, and atherosclerosis. IL-6 signals through a receptor complex of IL-6 R alpha and gp130. gp130 is also a component of the receptors for CLC, CNTF, CT-1, IL-11, IL-27, LIF, and OSM. Soluble forms of IL-6 R alpha are generated by both alternative splicing and proteolytic cleavage. In a mechanism known as trans-signaling, complexes of soluble IL-6 and IL-6 R alpha elicit responses from gp130-expressing cells that lack cell surface IL-6 R alpha.
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