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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
20 pg/mL
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Assay Range
39.0 - 2,500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Specificity
Natural and recombinant human G-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 G-CSF Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure G-CSF in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human G-CSF and antibodies raised against the protein. It has been shown to accurately quantitate recombinant human G-CSF. Results obtained using natural human G-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 human G-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 | 20 | 20 | 20 |
| Mean | 177 | 515 | 1047 | 143 | 818 | 1514 |
| Standard Deviation | 4.5 | 9.4 | 22 | 17 | 72 | 77.1 |
| CV% | 2.5 | 1.8 | 2.1 | 11.9 | 8.8 | 5.1 |
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 | 280 | 827 | 1696 | 176 | 1094 | 2169 |
| Standard Deviation | 7.8 | 14.1 | 19.5 | 7.3 | 34.6 | 81.6 |
| CV% | 2.8 | 1.7 | 1.1 | 4.1 | 3.2 | 3.8 |
Recovery
The recovery of G-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) | 94 | 85-110 |
| Citrate Plasma (n=5) | 90 | 78-111 |
| EDTA Plasma (n=5) | 93 | 80-117 |
| Heparin Plasma (n=5) | 87 | 76-103 |
| Serum (n=5) | 88 | 75-106 |
Linearity
To assess the linearity of the assay, five samples were spiked with high concentrations of G-CSF in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Background: G-CSF
G-CSF (granulocyte-colony stimulating factor) is a secreted glycoprotein that regulates the proliferation, differentiation, and activation of neutrophilic granulocyte lineage cells. It also enhances M-CSF induced monocyte differentiation and the proliferation of Th2-inducing dendritic cells. G-CSF promotes the development of T cell immune tolerance as well as tissue recovery following myocardial infarction and cerebral ischemia. It signals through G-CSF R/CD114 on monocytes, neutrophils, megakaryocytes, platelets, myeloid progenitors, trophoblasts and placenta, endothelial cells, and various tumor cell types. Mutations in G-CSF R are associated with congenital neutropenia.
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