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If the gradients are prepared the day before the experiment and left in the refrigerator overnight then this can be a convenient approach

Following 16 h incubation (at 4 C), the wells were washed with PBS/0.1% BSA and the bound radioactivity (representing 3 H-25HC bound to v3 integrin in the absence and presence of fractalkine-CD protein) was counted using a TRI-CARB 2900TR liquid scintillation counter

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Ho YS, Magnenat JL, Bronson RT, Cao J, Gargano M, Sugawara M, Funk CD