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Sample Type & Volume Required Per Well
Cell Culture Supernates (200 uL), Serum (200 uL), EDTA Plasma (200 uL), Heparin Plasma (200 uL), Citrate Plasma (200 uL)
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Sensitivity
10 pg/mL
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Assay Range
31.2 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Specificity
Natural and recombinant human GRO alpha
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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 Human CXCL1/GRO alpha Immunoassay is a 4.25-4.5 hour solid phase ELISA designed to measure human GRO alpha in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human GRO alpha and antibodies raised against the recombinant protein. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural human GRO alpha showed linear 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 human GRO alpha.
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 twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
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 | 61.4 | 251 | 736 | 62.8 | 252 | 734 |
| Standard Deviation | 2.2 | 6 | 20 | 3.6 | 11.9 | 38.9 |
| CV% | 3.6 | 2.4 | 2.7 | 5.7 | 4.7 | 5.3 |
Cell Culture Supernates | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 81.4 | 318 | 734 | 87.4 | 320 | 758 |
| Standard Deviation | 3.7 | 7.4 | 55 | 11.9 | 27.6 | 62.7 |
| CV% | 4.5 | 2.3 | 7.5 | 13.6 | 8.6 | 8.3 |
Recovery
The recovery of human GRO alpha spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Media (n=4) | 97 | 81-108 |
| Cell Culture Media (n=4) | 97 | 81-108 |
| Citrate Plasma (n=10) | 90 | 80-98 |
| Citrate Plasma (n=10) | 90 | 80-98 |
| EDTA Plasma (n=10) | 93 | 86-102 |
| EDTA Plasma (n=10) | 93 | 86-102 |
| Heparin Plasma (n=10) | 95 | 83-109 |
| Heparin Plasma (n=10) | 95 | 83-109 |
| Serum (n=10) | 98 | 87-108 |
| Serum (n=10) | 98 | 87-108 |
Linearity
To assess linearity of the assay, samples containing and/or spiked with high concentrations of human GRO alpha were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
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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