Important Notes:

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Secrets to successful tandem use:

  • Avoid extended exposure to light and extreme variations in temperature. Light will photobleach your tandem – protect your samples and antibody from light at all times. Do not freeze your tandem antibody conjugates, as this could potentially lead to denaturing of the donor protein.
  • Avoid ”overfixing“. We recommend that tandems be fixed for short-periods of time (less than 30 minutes) and then washed and resuspended in Cell Staining buffer for storage. For this purpose, we provide specially formulated FluoroFix™ buffer, for the fixation of tandem dyes.
  • Be aware of cross-beam excitation of your acceptor molecule when performing multi-laser flow cytometry with tandems. In most cases, this can be compensated. For example, the Cy5 acceptor in PE/Cy5 is known to be excitable by the red laser, so use caution in combination with APC.
  • Label your cells at 4°C or on ice. APC tandems may be susceptible to cell-mediated decoupling, so slowing down cell metabolism at low temperatures can help preserve the stability of the tandem.