Mouse/Rat RGM-C/Hemojuvelin Quantikine ELISA Kit Summary

Product Summary

The Quantikine Mouse/Rat RGM-C/Hemojuvelin Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse and rat RGM-C in cell culture supernates, tissue homogenates, serum, and plasma. It contains NS0-expressed recombinant mouse RGM-C and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural RGM-C showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural RGM-C.

Preparation and Storage

Background: RGM-C/Hemojuvelin

The repulsive guidance molecule (RGM) family of molecules contains three uniquely structured GPI-linked glycoproteins that participate in a variety of activities. Termed RGM-A, -B and -C, they are 40-45 kDa in size, show generally nonoverlapping fetal expression patterns, and serve as BMP-4 receptors. RGM-A and -C are plasma membrane bound, while RGM-B is generally microsomal in location. RGM-A and -B are associated with neurons, while RGM-C regulates skeletal muscle and hepatocyte function.

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.