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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 (100 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
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Sensitivity
0.14 ng/mL
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Assay Range
0.8 - 50 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Specificity
Natural and recombinant human IGFBP-3
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Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
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Interference
No significant interference observed with available related molecules.
Control Available
QC23, Quantikine Immunoassay Control Group 8 -
Product Summary
The Quantikine Human IGFBP-3 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human IGFBP-3 in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human IGFBP-3 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human IGFBP-3 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 naturally occurring human IGFBP-3.
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, EDTA Plasma, Heparin Plasma | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 4.13 | 12.7 | 31.1 | 4.06 | 11.8 | 28.5 |
| Standard Deviation | 0.2 | 0.64 | 0.71 | 0.22 | 0.76 | 2.27 |
| CV% | 4.8 | 5 | 2.3 | 5.4 | 6.4 | 8 |
Recovery
The recovery of IGFBP-3 spiked to three different levels throughout the range of the assay was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Media (n=4) | 94 | 89-99 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of IGFBP-3 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Background: IGFBP-3
The superfamily of insulin-like growth factor (IGF) binding proteins include the six high-affinity IGF binding proteins (IGFBP) and at least four additional low-affinity binding proteins referred to as IGFBP related proteins (IGFBP-rP). All IGFBP superfamily members are cysteine-rich proteins with conserved cysteine residues, which are clustered in the amino- and carboxy-terminal thirds of the molecule. IGFBPs modulate the biological activities of IGF proteins. Some IGFBPs may also have intrinsic bioactivity that is independent of their ability to bind IGF proteins. Post-translational modifications of IGFBP, including glycosylation, phosphorylation and proteolysis, have been shown to modify the affinities of the binding proteins to IGF. ALS (Acid Labile Subunit) is a liver-derived protein that exists in a ternary complex with Insulin-like Growth Factor (IGF)-binding Protein-3 (IGFBP-3) or IGFBP-5, and either IGF-I or IGF-II. ALS increases the half-life of IGF/IGFBP complexes in circulation.
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