| Assay Method Information | |
| | IL-21 ALPHALISA assay |
| Description: | IL-21 levels in cell supernatants and cell-free control samples were measured using the ALPHALISA assay kit from Perkin Elmer, following the manufacturer's instructions. The assay was run at room temperature in 384-well ALPHAPLATES (cat. 6005350; Boston, MA). Briefly, immediately after transferring the cell supernatant to the assay plate, anti-IL-21 acceptor beads were added with the CERTUS FLEX liquid handler (LEAP Technologies, Morrisville, NC). The plate was sealed, spun down and incubated for 30 minutes. After incubation, the seal was removed, anti-IL-21 biotinylated antibody was added with the CERTUS FLEX, and the plate was sealed again, spun down and incubated for 60 minutes. Next, the seal was removed, streptavidin-coated donor beads were added with the CERTUS FLEX, and the plate was sealed, spun down and incubated for another 60 minutes. Finally, the seal was removed and the plate was read using the ENVISION reader (Perkin Elmer, Boston, MA) set with an excitation filter of 680 nm and an emission filter of 615 nm. Two independent IL-21 standard dilution curves were run per plate with analyte provided in the ALPHALISA kit. |
| Affinity data for this assay | |
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