< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
No significant interference observed with available related molecules.
The Quantikine Mouse MCP-5 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse MCP-5 levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse MCP-5 and antibodies raised against the recombinant factor. Results obtained for naturally occurring mouse MCP-5 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 mouse MCP-5.
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
The recovery of mouse MCP-5 spiked to three levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Supernates (n=6)
EDTA Plasma (n=6)
To assess the linearity of the assay, six samples containing and/or spiked with high concentrations of mouse MCP-5 in each matrix were diluted with Calibrator Diluent and then assayed.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Mouse MCP-5, also known as CCL12, encodes a 104 amino acid (aa) precursor protein with a 22 aa hydrophobic signal sequence that is cleaved to generate a 82 aa residue mature protein. MCP-5 is expressed constitutively in the thymus and lymph nodes. Under inflammatory conditions, MCP-5 expression is also induced in activated macrophages and mast cells.
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
50 µL Assay Diluent
Add 50 µL of Assay Diluent to each well.
50 µL Standard, Control, or Sample
Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
100 µL Conjugate
Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
Aspirate and wash 5 times.
100 µL Substrate Solution
Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
100 µL Stop Solution
Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
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