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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
3.5 hours or 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
3 pg/mL
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Assay Range
7.8 - 500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Specificity
Natural and recombinant human 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.
Control Available
QC01-1, Quantikine Immunoassay Control Group 1 -
Product Summary
The Quantikine Human GM-CSF Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure GM-CSF levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human GM-CSF and antibodies raised against the recombinant factor. It has been shown to quantitate recombinant human GM-CSF accurately. Results obtained using natural human GM-CSF 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 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 | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 46.8 | 131 | 269 | 45.1 | 133 | 266 |
| Standard Deviation | 1.1 | 3.6 | 7.3 | 2.4 | 7 | 11.5 |
| CV% | 2.4 | 2.7 | 2.7 | 5.3 | 5.3 | 4.3 |
Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 54.7 | 156 | 321 | 53.1 | 157 | 317 |
| Standard Deviation | 1.1 | 3.2 | 4.8 | 3.3 | 9.1 | 15.4 |
| CV% | 2 | 2.1 | 1.5 | 6.2 | 5.8 | 4.9 |
Recovery
The recovery of GM-CSF 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) | 97 | 85-104 |
| Citrate Plasma (n=5) | 101 | 92-106 |
| EDTA Plasma (n=5) | 100 | 94-105 |
| Heparin Plasma (n=5) | 98 | 92-104 |
| Serum (n=5) | 98 | 93-103 |
Linearity
To assess the linearity of the assay, five samples were spiked with high concentrations of GM-CSF in various matrices and diluted with the appropriate Calibrator Diluent to produce values within the dynamic range of the assay.
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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