Human Chitinase 3-like 1 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human CHI3L1 Immunoassay is a 4.5 hour solid phase ELISA designed to measure CHI3L1 in cell culture supernates, serum, plasma, and urine. It contains NS0-expressed recombinant human CHI3L1 and antibodies raised against recombinant human CHI3L1 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural CHI3L1 showed dose response 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 CHI3L1.

Preparation and Storage

Background: Chitinase 3-like 1

Chitinase 3-like 1 (CHI3L1), also called BRP39 in mouse or YKL-40 in humans, is a 39 kDa glycoprotein member of the glycosyl hydrolase 18 family although it does not show chitotriosidase activity. CHI3L1 is expressed by articular chondrocytes, synovial cells, activated monocyte-derived macrophages, neutrophils, endothelial cells, vascular smooth muscle cells, and some cancer cells. CHI3L1 binds to chitin and heparins, and it enhances cell adhesion, proliferation and tumor angiogenesis. Circulating CHI3L1 is elevated during inflammation and connective tissue remodeling such as arthritis, chronic obstructive pulmonary disease, diabetes, cardiovascular disease, inflammatory bowel disease, and liver cirrhosis. It is frequently upregulated in glioblastoma, myxoid chondrosarcoma, melanoma and carcinomas of the breast, thyroid, colon, lung, kidney, and ovary.

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.