Mouse IL-5 Quantikine ELISA Kit

Assay Type	Solid Phase Sandwich ELISA
Format	96-well strip plate
Assay Length	4.5 hours
Sample Type & Volume Required Per Well	Cell Culture Supernates (50 uL), Serum (50 uL)
Sensitivity	7 pg/mL
Assay Range	15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum)
Specificity	Natural and recombinant mouse IL-5
Cross-reactivity	< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested
Interference	No significant interference observed with available related molecules.

Product Summary

The Quantikine Mouse IL-5 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse IL-5 in cell culture supernates and mouse serum. It contains Sf 21-expressed recombinant mouse IL-5 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant mouse IL-5. Results obtained using natural mouse IL-5 showed linear curves that were parallel to the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse IL-5.

Preparation and Storage

Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage	Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: IL-5

IL-5 (Interleukin-5) is a secreted cytokine that is expressed as a covalent dimer. It is produced by activated Th2 cells, eosinophils, mast cells, and iNKT cells and promotes eosinophilic and basophilic differentiation and activation. IL-5 signals through a receptor complex containing IL-5 R alpha and the Common beta Chain/CD131 on eosinophils and basophils. Naturally occurring soluble forms of IL-5 R alpha retain the ability to bind IL-5 and limit its bioavailability.

Long Name:	Interleukin 5
Entrez Gene IDs:	3567 (Human); 16191 (Mouse); 24497 (Rat); 397409 (Porcine); 280825 (Bovine); 403790 (Canine); 493803 (Feline)
Alternate Names:	BCDF mu; B-cell differentiation factor I; BCGFII; EDF; Eo-CSF; Eosinophil differentiation factor; IL5; IL-5; IL-5T-cell replacing factor; interleukin 5 (colony-stimulating factor, eosinophil); interleukin-5; TRF; TRFB cell differentiation factor I

Assay Procedure

Refer to the product datasheet for the 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.

1. Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3. Add 50 μL of Assay Diluent to each well.

4. Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5. Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6. Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7. Aspirate and wash 5 times.

8. Add 100 μL Substrate Solution to each well.

9. 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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