Mouse CCL12/MCP-5 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Mouse MCP-5 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse MCP-5 levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse MCP-5 and antibodies raised against the recombinant factor. Results obtained for naturally occurring mouse MCP-5 showed linear curves that were parallel to the standard curves obtained using the kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse MCP-5.

Preparation and Storage

Background: CCL12/MCP-5

Mouse MCP-5, also known as CCL12, encodes a 104 amino acid (aa) precursor protein with a 22 aa hydrophobic signal sequence that is cleaved to generate a 82 aa residue mature protein. MCP-5 is expressed constitutively in the thymus and lymph nodes. Under inflammatory conditions, MCP-5 expression is also induced in activated macrophages and mast cells.

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.