Mouse Osteoprotegerin/TNFRSF11B Quantikine ELISA Kit Summary

Product Summary

The Quantikine Mouse OPG Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse OPG in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse OPG and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant mouse OPG accurately. Results obtained using natural mouse OPG showed dose response 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 mouse OPG.

Preparation and Storage

Background: Osteoprotegerin/TNFRSF11B

Osteoprotegerin (OPG), also known as OCIF and TNFRSF11B, is a secreted TNF superfamily receptor that regulates bone density. OPG is widely expressed and constitutively released as a homodimer by mesenchymal stem cells, fibroblasts, and endothelial cells. OPG functions as a decoy receptor for TRANCE/RANK L and TRAIL. OPG regulates the cell surface expression of TRANCE and its ability to stimulate osteoclastogenesis. Dysregulation of OPG/TRANCE interactions can lead to juvenile Paget’s disease, osteoporosis, bone loss, and vascular calcification..

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.