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Description of Methods for the Use of MONOSAN® Antibodies in Indirect Flow Cytometry

Surface Staining:

  1. Prepare cell suspension in PBS.
  2. Add 10µl of antibody at various dilutions eg 1:10 upwards to 90µl of cell suspension. Incubate for 30 minutes at 4°C (an isotype control is recommended).
  3. Wash twice in PBS.
  4. Add Secondary FITC Ab (or equivalent) in PBS. Incubate for 30 minutes at 4°C.
  5. Wash twice in PBS.
  6. Resuspend in 300µl of PBS for analysis.

Intracellular Staining:

  1. Prepare cell suspension in PBS (containing 0.5% (w/v) saponin).
  2. Add 10µl of antibody at various dilutions eg 1:10 upwards to 90µl of cell suspension. Incubate for 30 minutes at 4°C (an isotype control is recommended).
  3. Wash twice in PBS.
  4. Add secondary FITC Ab (or equivalent) in PBS. Incubate for 30 minutes at 4°C.
  5. Wash twice with PBS.
  6. Resuspend in 300µl of PBS for analysis.

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