Human Resistin Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human Resistin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Resistin in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human Resistin and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Resistin 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 naturally occurring Resistin.

Preparation and Storage

Background: Resistin

Resistin, also known ADSF and FIZZ3, is a secreted peptide hormone that circulates as either a noncovalent trimer, or a disulfide-linked homohexamer. Resistin is expressed by white adipocytes, splenocytes, astrocytes, and anterior pituitary epithelium. It blocks Insulin-stimulated uptake of glucose by adipocytes and promotes glucose release by hepatocytes. High circulating Insulin levels induce Resistin secretion which then antagonize Insulin action.

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.