Question: Do erythrocytes (red blood cells) interfere with the assay?
Answer: The large number of erythrocytes present in peripheral blood, apheresis collections, bone marrow, and umbilical cord blood samples can compete with stem/progenitor cells for the ALDEFLUOR® substrate. Ammonium chloride treatment of the samples to lyse the erythrocytes, gives optimal assay performance. The ratio of lysis buffer to cell numbers or blood volume must be optimized (10 to 40 parts buffer to sample), and the time (10 - 30 minutes) and temperature (RT or 2 - 8°C) of incubation must be carefully controlled for each lysis buffer and sample type.
Question: Can I run the staining reaction at room temperature (15 - 25°C)?
Answer: Yes. However, the staining reaction may take 2 - 4 hours to reach completion, depending on the temperature of the room.