The capture beads are bottled individually, and it is necessary to pool the six individual bead reagents immediately before mixing them together with the PE-detection reagent, standards, and samples.
1. Determine the number of assay tubes (including standards and controls) that are required for the experiment (e.g., 8 unknowns, 9 cytokine standard dilutions, and 1 negative control = 18 assay tubes).
2. Vigorously vortex each capture bead suspension for a few seconds before mixing.
3. Add a 10-p.L aliquot of each capture bead, for each assay tube to be analyzed, into a single tube labeled "mixed capture beads" (e.g., 10 ¡L of IL-2 capture beads x 18 assay tubes = 180 ¡L of IL-2 capture beads required).
4. Vortex the bead mixture thoroughly. The mixed capture beads are now ready to be transferred to the assay tubes.
Was this article helpful?