Specimen and Reagent Preparation for Quantitative Flow Cytometry

1. To quantitate CRKL, label four 12 X 75-mm flow tubes—one tube each for total and phosphorylated primary CRKL antibodies and one tube each for each primary antibody's isotype controls (see Note 2).

2. Pipet the appropriate volume of peripheral blood (equivalent to 1 X 106 cells) into each of the four flow tubes and incubate in the presence of either 10 p,L surface markers or 5 p,L isotype controls for each fluorochrome for 15 min at room temperature in the dark.

M2 M3 M4

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