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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), Tissue Homogenates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
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Sensitivity
3 pg/mL
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Assay Range
7.8 - 500 pg/mL (Cell Culture Supernates, Tissue Homogenates, Serum, EDTA Plasma)
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Specificity
This kit recognizes both the 164 and 120 amino acid residue forms of mouse VEGF
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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
Interference observed with 1 or more available related molecules.
Product Summary
The Quantikine Mouse VEGF Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse VEGF in cell culture supernates, tissue homogenates, mouse serum, and plasma. It contains Sf 21-expressed mouse VEGF and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse VEGF accurately. Results obtained using natural mouse VEGF showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse VEGF.
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, Tissue Homogenates, Serum, EDTA Plasma | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 37.7 | 144 | 277 | 38.3 | 138 | 291 |
| Standard Deviation | 3.1 | 6.2 | 12.9 | 3.2 | 7.8 | 18.5 |
| CV% | 8.2 | 4.3 | 4.7 | 8.4 | 5.7 | 6.4 |
Recovery
The recovery of mouse VEGF spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Supernates (n=9) | 103 | 95-114 |
| EDTA Plasma (n=5) | 91 | 82-103 |
| Heparin Plasma (n=4) | 89 | 87-94 |
| Serum (n=9) | 101 | 83-115 |
| Tissue Homogenates (n=3) | 91 | 76-107 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with various concentrations of mouse VEGF in each matrix were diluted with Calibrator Diluent and then assayed.
Preparation and Storage
Background: VEGF
VEGF (Vascular endothelial growth factor) is a potent mediator of both angiogenesis and vasculogenesis in the fetus and adult. It is required for regulating the proliferation, migration, and survival of embryonic endothelial cells and during wound healing and the female reproductive cycle in adults. Pathologically, it is involved in tumor development and tumor vascular leakage. VEGF binds to VEGF R1/Flt-1, VEGF R2/Flk-1/KDR, and Neuropilin-1. Multiple forms of VEGF are generated by alternative splicing and proteolysis.
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