Mouse/Rat WISP-1/CCN4 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Mouse/Rat WISP-1/CCN4 immunoassay is a 4.5 hour solid-phase ELISA designed to measure WISP-1 in mouse or rat cell culture supernates, tissue lysates, serum, plasma, and urine. It contains NS0-expressed recombinant mouse WISP-1 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant mouse WISP-1. Results obtained using natural mouse or rat WISP-1 showed dose response curves that were parallel to the standard curves obtained using the Quantikine mouse kit standards. These results indicate that this kit can be used to determine relative mass values for natural WISP-1.

Preparation and Storage

Background: WISP-1/CCN4

WISP-1 belongs to the CCN (CTGF/CYR61/NOV) family of secreted proteins that share a common multimodular organization. Each family member has an IGFBP domain, a von Willebrand type C domain, a thrombospondin 1 domain and a C-terminal cysteine knot domain. Depending on cell context, WISP-1 can either promote or inhibit tumorigenesis.

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.