Mouse/Rat FGF-21 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Mouse/Rat FGF-21 immunoassay is a 4.5 hour solid-phase ELISA designed to measure FGF-21 in mouse or rat cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse FGF-21 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant mouse FGF-21. Results obtained using natural mouse or rat FGF-21 showed dose response curves that were parallel to the standard curves obtained using the Quantikine mouse kit standards. These results indicate that this kit can be used to determine relative mass values for natural FGF-21.

Preparation and Storage

Background:  FGF-21

FGF-21 (Fibroblast Growth Factor 21) is a member of an FGF subfamily that also includes FGF-19 and FGF-23. FGF-19 subfamily members lack a heparin binding domain, enabling them to freely circulate as endocrine factors and diffuse within tissues. FGF-21 signals through Klotho beta in complex with FGF receptors. It regulates cellular metabolism including glucose uptake in adipocytes and cellular sensitivity to insulin. It is basally expressed in the pancreas, thymus, liver, and adipose tissue and is upregulated in liver, muscle, and fat in obesity and diabetes. FGF-21 also plays a role in cell survival and proliferation, mesenchymal stem cell differentiation, circadian rhythm, and controlling reproductive capacity.

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.