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Assay Length
4.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL)
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Sensitivity
2 pg/mL
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Assay Range
15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum)
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Specificity
Natural and recombinant mouse KC
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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 Mouse KC Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse KC in cell culture supernates and mouse serum. It contains E. coli-expressed recombinant mouse KC and antibodies raised against the recombinant factor. This Immunoassay has been shown to quantitate the recombinant mouse KC accurately. Results obtained using natural mouse KC showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse KC.
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 | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 34.9 | 169 | 530 | 35 | 183 | 521 |
| Standard Deviation | 1.9 | 5.2 | 26.3 | 2.1 | 18 | 15.6 |
| CV% | 5.4 | 3.1 | 5 | 6 | 9.8 | 3 |
Recovery
The recovery of mouse KC spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Supernates (n=7) | 98 | 89-103 |
| Serum (n=9) | 95 | 83-104 |
Linearity
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse KC in each matrix were diluted with Calibrator Diluent and then assayed.
Preparation and Storage
Background: CXCL1/GRO alpha/KC/CINC-1
CXCL1/GRO alpha, also known as KC in mouse and CINC-1 in rat, is a member of the CXC family of chemokines. Human CXCL1/GRO alpha is 107 amino acids (aa) in length with a predicted molecular weight of 11 kDa. The mouse and rat orthologs share 57% and 59% amino acid sequence identity with the human protein, respectively. Genome-wide analysis suggests that the protein encoded by the mouse Cxcl3 gene is homologous to human CXCL1/GRO alpha protein. Human CXCL1/GRO alpha can be cleaved into three isoforms CXCL1/GRO alpha (aa 4-73), CXCL1/GRO alpha (aa 5-73), and CXCL1/GRO alpha (aa 6-73) which show higher activity than the full length protein. CXCL1/GRO alpha is secreted by mast cells and macrophages whereupon it acts as a chemoattractant for neutrophils.
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