How do we perform FasL lab test?
Protocols we follow to perform FasL testing
We, at NorthEast BioLabs, use MSD, ELISA, and Luminex assays to detect FasL protein in human serum, plasma, and cell culture supernatants. Below, we explain a Sandwich-based ELISA protocol for detecting Fas ligand protein in human samples.
We begin with adding an capture antibody to coat the microplate by incubating them for 2.5 hrs at room temperature (RT) or O/N at 4℃. After this incubation, we block the plates with 1% bovine serum albumin or skimmed milk for 1 hr at RT. Next with 100 μl of serially diluted standards and samples are added to respective wells and incubate them for 2.5 hrs at room temperature (RT). We then wash the plate using a plate washer and introduce a biotinylated antibody in each well and incubate it for 1 hr at RT. Then 100 μl of streptavidin-conjugated HRP is added to each well and incubated for 45 mins at RT. We then incubate the reaction mixture with TMB one-step substrate reagent for 30 min at RT. Lastly, we add 50 μl of stop solution and read the reaction at 450 nm.
The Fas ligand assay is a quantitative, colorimetric-based assay for the detection of FasL. The assay has 2 pg/ml sensitivity and can detect between 2 pg/ml to 1000 pg/ml.