Mouse MFG-E8 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Mouse MFG-E8 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse MFG-E8 in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse MFG-E8 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural mouse MFG-E8 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 mouse MFG-E8.

Preparation and Storage

Background: MFG-E8

MFG-E8, originally identified as a product of lactating mammary glands, is also secreted by activated macrophages and immature dendritic cells. MFG-E8 functions as an opsonin by binding phosphatidylserine on apoptotic cells and enhancing their phagocytosis.

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.