Preparation of Soluble Protein Flex Set Capture Beads

The capture beads provided with each soluble protein Flex Set are a 50X bulk (1 ^L/test) and should be mixed with other soluble protein Flex Set capture beads and diluted to their optimal volume per test (50 ^L/test) before adding the beads to a given tube or assay well.

1. Determine the number of soluble protein Flex Sets to be used in each tube or assay well in the experiment (size of the multiplex).

2. Determine the number of tubes or wells in the experiment.

3. Vortex each Soluble Protein Flex Set capture bead and then transfer 1 ^L/test of each capture bead to a conical tube labeled "mixed capture beads."

a. If testing cell culture supernatant samples, add capture bead diluent to the mixed capture beads tube to bring the final volume to 50 ^L/test.

b. If testing serum or plasma samples, add 0.5 mL of wash buffer to the mixed capture beads tube and centrifuge at 200g for 5 min to pellet the beads. Discard the supernatant by aspiration. Resuspend beads in capture bead diluent for serum/plasma to a final volume of 50 ^L/test and incubate for 15 min at room temperature before proceeding to step 5.

Example: if five soluble protein Flex Sets are being multiplexed for a given 20 test experiment, you would add 1 pL/test of each capture bead to the mixed capture Bead tube (1 pL/test X 20 tests = 20 pL total volume of each soluble protein Flex Set capture bead) and then add capture bead diluent to bring the final volume to 50 pL/test by determining the remaining volume to add (the final volume of mixed capture beads is 20 tests X 50 pL/test = 1000 pL). A total of 100 pL of capture beads were added to the mixed capture beads tube previously listed when 20 pL total volume of each capture bead was added from the five soluble protein Flex Sets. The amount of capture bead diluent to add is 1000 pL total volume - 100 pL of capture beads = 900 pL).

5. Vortex the beads to mix thoroughly. Mixed capture beads are now ready to be used in the experiment.

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