Human IL-7 Quantikine HS ELISA Kit Summary

Product Summary

The Quantikine HS Human IL-7 Immunoassay is a solid phase ELISA that specifically measures IL-7 in 18-24 hours (with an overnight incubation step). This kit is designed to measure IL-7 levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human IL-7 and antibodies raised against the recombinant factor. It has been shown to quantitate the recombinant factor accurately. Results obtained using natural IL-7 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 human IL-7.

Preparation and Storage

Background: IL-7

IL-7 (Interleukin-7) is a cytokine that plays important roles in lymphocyte differentiation, proliferation, and survival. IL-7 is produced by stromal epithelial cells of the thymus, bone marrow, and intestines. It signals through a receptor complex composed of IL-7 R alpha/CD127 and the Common gamma Chain. gamma c is also a subunit of the receptors for IL-2, -4, -9, -15, and -21. IL-7 contributes to the maintenance of all naïve and memory T cells. It is required for optimal T cell-dendritic cell interaction. In mouse, IL-7 activation of IL-7 R alpha is critical for both T cell and B cell lineage development, while in humans, it is required for T cell but not for B cell development.

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.