Preparation of Mixed Human ThTh Cytokine Capture Beads

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.

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