Human CCL7/MCP-3 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human MCP-3 Immunoassay is a 4.5 hour solid phase ELISA designed to measure MCP-3 levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human MCP-3 and antibodies raised against the recombinant factor. Results obtained for naturally occurring human MCP-3 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 human MCP-3.

Preparation and Storage

Background: CCL7/MCP-3/MARC

Human MCP-3 and mouse MARC are members of the CC subfamily of chemokines and are monocyte chemoattractants. Sequence comparisons suggest that MARC may be the mouse homolog of the human MCP-3 gene. Except for one amino acid subsititution, mouse MARC is identical to mouse FIC, the product of a growth factor-activated gene.

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.