Mouse Adiponectin/Acrp30 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Mouse Adiponectin Immunoassay is a 4.5 hour solid phase ELISA designed to measure full-length mouse Adiponectin levels in cell culture supernates, tissue homogenates, serum, and plasma. It contains NS0-expressed recombinant mouse Adiponectin and antibodies raised against the recombinant protein. Results obtained for naturally occurring mouse Adiponectin 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 of natural mouse Adiponectin.

Preparation and Storage

Background: Adiponectin/Acrp30

Adiponectin (also known as Adipocyte complement-related protein of 30 kDa, Acrp30) is an adipocyte-derived protein with wide ranging paracrine and endocrine effects on metabolism and inflammation. It promotes adipocyte differentiation, fatty acid catabolism, and insulin sensitivity and is negatively correlated with obesity, type 2 diabetes, and atherogenesis. Adiponectin associates into 70 kDa noncovalently-linked trimers that are disulfide-linked into medium molecular weight hexamers and then into >300 kDa high molecular weight oligomers.

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.