Human MIA Quantikine ELISA Kit

Product Summary

The Quantikine Human MIA Immunoassay is a 4.5 hour solid-phase ELISA designed to measure MIA in cell culture supernates, serum, plasma, and saliva. It contains E. coli-expressed recombinant Human MIA and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant human MIA. Results obtained using natural human MIA 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 natural human MIA.

Preparation and Storage

Background: MIA

MIA, also known as cartilage-derived retinoic acid-sensitive protein (CD-RAP), is a secreted protein expressed in cartilaginous tissue during development and in the adult. MIA was identified based on the fact that it is highly expressed in malignant melanomas, but not in normal melanocytes. MIA has been shown to interact with fibronectin, and promotes invasion and metastasis of melanoma cells, perhaps by blocking integrin-fibronectin interactions.

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.