PE Mouse Anti-Human CD135

Protocol tips

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	Ab incubation at room temperature in the dark. Red blood cells were then lysed with NH4CL (0.83%) at 37°C for 5 minutes before undergoing centrifugation. Cells were then washed in buffered saline and re-suspended in 1% paraformaldehyde analyses on a FACSCalibur (Becton Dickinson Biosciences) and 200,000 events were collected for each tube.

Protocol tips
Ab incubation at room temperature in the dark. Red blood cells were then lysed with NH4CL (0.83%) at 37°C for 5 minutes before undergoing centrifugation. Cells were then washed in buffered saline and re-suspended in 1% paraformaldehyde analyses on a FACSCalibur (Becton Dickinson Biosciences) and 200,000 events were collected for each tube.

Publication protocol

CD135 mean fluorescent intensity (MFI) of the myeloid blast population, as defined by CD45 low and side scatter, was determined by multi-dimensional flow cytometry (MDF). Specimens were processed as previously described.(22) Bone marrow cells were incubated with CD34-APC and CD45-PerCP (BD Biosciences, San Jose, CA), CD135- PE) (BD Pharmingen, San Diego, CA), and C36-flourescein (FITC) (Beckman Coulter, Brea, CA) at room temperature in the dark. Red blood cells were then lysed with NH4CL (0.83%) at 37°C for 5 minutes before undergoing centrifugation. Cells were then washed in buffered saline and re-suspended in 1% paraformaldehyde analyses on a FACSCalibur (Becton Dickinson Biosciences) and 200,000 events were collected for each tube. Linear MFI was determined as previously described.(23)

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Manufacturer protocol

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