Mouse TRANCE/RANK L/TNFSF11 Quantikine ELISA Kit Summary

Product Summary

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

Preparation and Storage

Background: TRANCE/TNFSF11/RANK L

TRANCE (TNF-related activation-induced cytokines), also known as RANK Ligand, OPGL, or ODF, is a TNF superfamily ligand produced by osteoblasts, bone marrow stromal cells, and activated T cells. It signals through RANK and is antagonized by the decoy receptor Osteoprotegerin. TRANCE stimulates bone resorption, is required for osteoclast differentiation, and is associated with the development of osteoporosis, vascular calcification, osteopetrosis, and progestin-induced breast cancer. It also mediates body temperature regulation and is essential for the development of a lactating mammary gland.

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.