Human Fas/TNFRSF6/CD95 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human sFas Immunoassay is a 4.5 hour solid phase ELISA designed to measure human sFas in cell culture supernates, serum, and plasma. It contains recombinant human Fas/Fc chimera expressed from NS0 cells and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained measuring natural human sFas showed dose-response curves that were parallel to the standard curves obtained using the recombinant Fas/Fc chimera. These results indicate that this kit can be used to determine relative mass values for natural human sFas.

Preparation and Storage

Background: Fas/TNFRSF6/CD95

Fas (Fibroblast-associated), also known as Apo-1, CD95, and TNFRSF6, was originally identified as a cell-surface protein that binds to monoclonal antibodies that are cytolytic for various human cell lines. Alternatively spliced cDNAs encoding multiple Fas isoforms, including a soluble form of Fas, have been identified. Fas is highly expressed in epithelial cells, hepatocyes, activated mature lymphocytes, virus-transformed lymphocytes and other tumor cells. Fas expression has also been detected in mouse thymus, liver, heart, lung, kidney and ovary.

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.