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AlphaScreen SureFire Bead Titration Protocol

Below is an example of how to make your master mixes to perform a bead titration for most 2-step kits (where the original protocol recommends mixing the Reaction buffer, Activation buffer, and Acceptor beads in a 40:10:1 ratio). This ratio is indicated in the manual, in the "Buffer preparations and Storage Conditions" table. For other kits, you can titrate the beads similarly by reducing only the volume of beads in the mix (by a factor of 2 or 4).

SureFire bead titration

Though the final volume of the mixes (and therefore the concentrations of reaction buffer, activation buffer, dilution buffer, and Donor and Acceptor beads) changes slightly, we recommend that you perform the bead titration experiments by altering the volume of the master mixes in this manner.