How do we perform cystatin C lab test?
Protocols we follow to perform Cystatin C tests
At NorthEast BioLabs, we perform MSD, ELISA, and Luminex assays for testing cystatin C in biological and chemical samples. Below is a detailed Luminex assay protocol followed for a cystatin c test.
To assay for cystatin c , we begin with 200 μl of assay buffer in each well, shake it for 10 mins at RT, and decant it. Once we remove all the residues, we add 25 μl of the following components in appropriate wells: standard or control followed by assay buffer, matrix solution, diluted samples, and beads and are incubated overnight at 4 ℃. Once the beads capture an analyte, we add 50 μl of biotinylated detection antibody and incubate it for 1 hr at RT. We then incubate the wells with a 50 μl streptavidin-PE conjugate, a reporter molecule. Lastly, we add 100 μl of the sheath or drive fluid and read the plate on a Luminex platform.
The assay tests the cystatin c levels and identifies each microsphere through a fluorescent reporter signal system. The assay is sensitive (0.16 ng/ml – 0.24ng/ml) with an intra-assay precision of < 10% and inter-assay precision of < 20%. Moreover, the assay is highly accurate with 95% recovery (overnight protocol) and 105% recovery (2 hr protocol) in the serum matrix.