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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 (50 uL), Serum (50 uL)
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Sensitivity
5.8 pg/mL
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Assay Range
7.8 - 500 pg/mL (Cell Culture Supernates, Serum)
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Specificity
Natural and recombinant mouse GM-CSF
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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 GM-CSF Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse GM-CSF in cell culture supernates and serum. It contains E. coli-expressed recombinant mouse GM-CSF and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse GM-CSF accurately. Results obtained using natural mouse GM-CSF showed dose-response curves that were parallel to the standard curves obtained using the recombinant standards. These results indicate that this kit can be used to determine relative mass values for natural mouse GM-CSF.
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, Plasma | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 18.5 | 67.2 | 219 | 20 | 71.7 | 243 |
| Standard Deviation | 0.7 | 2.1 | 6.9 | 1 | 3.8 | 11.2 |
| CV% | 3.8 | 3.1 | 3.2 | 5 | 5.3 | 4.6 |
Recovery
The recovery of mouse GM-CSF spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Supernates (n=8) | 96 | 82-114 |
| Serum (n=7) | 94 | 82-112 |
Linearity
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse GM-CSF in each matrix were diluted with Calibrator Diluent and then assayed.
Preparation and Storage
Background: GM-CSF
GM-CSF (Granulocyte-Macrophage Colony Stimulating Factor) induces the development of monocytes, neutrophils, eosinophils, and myeloid and dermal dendritic cells. It also acts as a neutrophil and dendritic cell chemoattractant. GM-CSF promotes Th1 and Th17 cell mediated autoimmune inflammation as well as the inflammatory activation of dendritic cells, microglia, alveolar macrophages, and eosinophils. In addition, it cooperates with G-CSF in promoting tumor cell proliferation and invasion. GM-CSF signals through a receptor complex composed of GM-CSF R alpha and the Common beta Chain (beta c) with Syndecan-2 as a potential co-receptor. The beta c subunit is shared by the receptor complexes for IL-3 and IL-5.
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