Preparation of Capture Beads

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

1. Determine the number of BD CBA cell signaling Flex Sets to be used in each tube or assay well in the experiment (size of the multiplex).

2. Determine the number of assay of tubes or wells to be run in the experiment.

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

4. Add capture bead diluent to the mixed capture beads tube to bring the final volume to 50 ¡L/test.

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

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

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