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Propidium Iodide or 7­-AAD Viability Staining Protocol

Posted by Michaela Frister on Aug 21, 2014 10:23:56 AM

Propidium Iodide (PI) Cat. No. 13­-6990

7-­AAD Cat. No. 13-­6993

Note: Propidium Iodide and 7-­AAD are not compatible with intracellular staining protocols that require fixation and permeabilization. For these applications we recommend use of Tonbo Biosciences Ghost Dyes.

Caution: Propidium Iodide and 7­-AAD are potential carcinogens. Protect skin and eyes by wearing suitable protective clothing, gloves and eye/face protection.

Other Materials Required

  • Flow Cytometry Staining Buffer (Stain Buffer) (1X PBS with 2% FBS, 0.09% Na­-Azide)

Experimental Procedure

  1. Stain cell surface antigen(s).
  2. Wash cells in Stain Buffer. Centrifuge at 300-­400 x g for 5 minutes at room temperature and discard supernatant.
  3. Resuspend in appropriate volume of Stain Buffer for acquisition, typically 0.5­-1 mL.
  4. For PI: add 10 uL (0.5 µg) to each sample.
    1. OR
  5. For 7-­AAD: add 5 uL (0.25 µg) to each sample.
  6. Incubate samples for 5­-10 minutes in the dark prior to data acquisition.
  7. Do not wash the cells after addition of Propidium Iodide or 7-­AAD.
  8. Analyze cells as soon as possible after the incubation period as prolonged exposure to Propidium Iodide or 7­-AAD can have adverse effects on cell viability.

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