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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), Serum (50 uL)
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Sensitivity
1.5 pg/mL
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Assay Range
7.8 - 500 pg/mL (Cell Culture Supernates, Serum)
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Specificity
Natural and recombinant mouse MIP-2
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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.
Product Summary
The Quantikine Mouse MIP-2 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse MIP-2 levels in cell culture supernates and serum. It contains E. coli-expressed recombinant mouse MIP-2 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse MIP-2 accurately. Results obtained using natural mouse MIP-2 showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse MIP-2.
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 | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 17.7 | 53.9 | 363 | 16.9 | 52.1 | 346 |
| Standard Deviation | 0.5 | 1.3 | 7.3 | 1 | 2.3 | 15.4 |
| CV% | 2.8 | 2.4 | 2 | 5.9 | 4.4 | 4.5 |
Recovery
The recovery of mouse MIP-2 spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Supernates (n=10) | 96 | 84-110 |
| Serum (n=14) | 100 | 94-116 |
Linearity
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse MIP-2 in each matrix were diluted with Calibrator Diluent and then assayed.
Preparation and Storage
Background: CXCL2/GRO beta/MIP-2/CINC-3
CXCL2/GRO beta, also called MIP-2 in mouse and CINC-3 in rat, is a member of the CXC chemokine family. Human CXCL2/GRO beta is 107 amino acids (aa) in length with a predicted molecular weight of 11 kDa. The mouse and rat orthologs share 70% and 71% aa sequence identity with the human protein, respectively. N-terminal aa 1-4 of CXCL2/GRO beta can be post-translationally cleaved which confers enhanced hematopoietic bioactivity. CXCL2/GRO beta is produced by a variety of cell types including monocytes and macrophages at sites of inflammation and is chemotactic for granulocytes, including neutrophils.
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