Human IFN-beta Quantikine ELISA Kit Summary

Product Summary

The Quantikine® Human IFN-beta Immunoassay is a 4.5 hour solid phase ELISA designed to measure human IFN-beta in cell culture supernates. It contains CHO cell-expressed recombinant human IFN-beta and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human IFN-beta showed linear 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 natural human IFN-beta.

Preparation and Storage

Background: IFN-beta

Interferon-beta (IFN-beta) is a cytokine that is produced by fibroblasts and pathogen-exposed dendritic cells, macrophages, and endothelial cells. It signals through the heterodimeric IFN-alpha/beta Receptor. IFN-beta-deficient mice show increased susceptibility to experimental autoimmune encephalomyelitis (EAE), a disease model of human multiple sclerosis (MS). Furthermore, IFN-beta has been shown to suppress the Th17 cell response in both MS and EAE and has commonly been used as a treatment for MS.

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.