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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 (200 uL), Serum (10 uL), EDTA Plasma (200 uL), Heparin Plasma (200 uL), Platelet-poor Citrate Plasma (200 uL), Urine (10 uL)
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Sensitivity
0.7 pg/mL
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Assay Range
3.9 - 250 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Platelet-poor Citrate Plasma, Urine)
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Specificity
Natural and recombinant human EGF
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Cross-reactivity
Cross-reactivity observed with 1 or more 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
QC21, Quantikine Immunoassay Control Group 4 -
Product Summary
The Quantikine Human EGF Immunoassay kit is a 3.5 or 4.5 hour solid phase ELISA designed to measure EGF levels in cell culture supernates, serum, plasma, and urine. It contains E. coli-derived recombinant human EGF and antibodies raised against the recombinant factor. It has been shown to accurately quantitate recombinant human EGF. Results obtained measuring natural human EGF showed linear curves that were parallel to the standard curves obtained using the kit standards. These results indicate that this kit can be used to determine relative mass values for natural EGF.
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.
Serum, EDTA Plasma, Heparin Plasma, Platelet-poor Citrate Plasma | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 13.5 | 80 | 133 | 14.7 | 77 | 134 |
| Standard Deviation | 0.6 | 2 | 2.7 | 0.9 | 3.3 | 6.1 |
| CV% | 4.4 | 2.5 | 2 | 6.1 | 4.3 | 4.6 |
Cell Culture Supernates, Urine | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 15 | 78.9 | 134 | 15 | 80 | 136 |
| Standard Deviation | 0.4 | 2.5 | 4 | 0.9 | 4.5 | 6.8 |
| CV% | 2.7 | 3.2 | 3 | 6 | 5.6 | 5 |
Recovery
The recovery of EGF spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Media (n=5) | 98 | 88-111 |
| Citrate Plasma (n=10) | 104 | 89-115 |
| EDTA Plasma (n=10) | 100 | 80-111 |
| Heparin Plasma (n=10) | 96 | 79-108 |
| Serum (n=10) | 97 | 80-106 |
| Urine (n=1) | 102 | 97-108 |
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of EGF 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: EGF
EGF (Epidermal Growth Factor) is expressed as a transmembrane protein that is proteolytically cleaved to generate soluble forms. It signals through EGF R/ErbB1 which heterodimerizes with ErbB2, ErbB3, or ErbB4 and can also associate with PDGF R beta and HGF R/c-MET. During development, EGF regulates thymocyte differentiation, neuroglia production, and adipocyte maturation. In the adult, EGF plays a role in mammary gland lactogenesis, fibroblast mitosis, dissociation of the extracellular matrix, and cell migration.
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Long Name
Epidermal Growth Factor
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