< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.
The Quantikine Mouse IL-3 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse IL-3 in cell culture supernates and mouse serum. It contains E. coli-expressed recombinant mouse IL-3 and antibodies raised against recombinant mouse IL-3. This Immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse IL-3 showed dose 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 mouse IL-3.
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
The recovery of mouse IL-3 spiked to three levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Supernates (n=5)
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse IL-3 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.
Interleukin-3 (IL-3) is a secreted cytokine that stimulates the proliferation and differentiation of pluripotent hematopoietic stem cells and various lineage-committed progenitors. It also regulates the activation of mature mast cells, basophils, eosinophils, and macrophages. It is produced by activated T cells, thymic epithelial cells, activated mast cells, keratinocytes, neurons, and astrocytes. IL-3 signals through a receptor complex that contains IL-3 R alpha/CD123 and the common beta chain which is also a component of the receptors for IL-5 and GM-CSF. Receptors for IL-3 are present on bone marrow progenitors, macrophages, mast cells, eosinophils, megakaryocytes, basophils, and various myeloid leukemic cells. Human IL-3 does not show activity on mouse 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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