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Assay Length
4.5 hours
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Specificity
Natural and recombinant rat 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.
Product Summary
The Quantikine Rat IL-6 Immunoassay is a 4.5 hour solid phase ELISA designed to measure rat IL-6 in cell culture supernates, rat serum, and plasma. It contains E. coli-expressed recombinant rat IL-6 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant protein. Results obtained using natural rat IL-6 showed dose-response curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural rat 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, Serum, EDTA Plasma | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 22 | 27 | 28 |
| Mean | 120 | 256 | 1028 | 134 | 268 | 987 |
| Standard Deviation | 10.6 | 22.4 | 46.2 | 13.4 | 21.7 | 69.2 |
| CV% | 8.8 | 8.8 | 4.5 | 10 | 8.1 | 7 |
Recovery
The recovery of rat IL-6 spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Supernates (n=5) | 103 | 85-118 |
| EDTA Plasma (n=5) | 96 | 81-111 |
| Serum (n=5) | 97 | 91-105 |
Linearity
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of rat IL-6 in each matrix were diluted with Calibrator Diluent and then assayed.
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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