RITs were labeled with the Alexa Fluor 647 Labeling Kit (Invitrogen) for 3.5 hrs and purified according to manufacturer’s instructions. Harvested cells were incubated for 30, 60 and 120 min at 37°C with 2 μg/ml of SS1P-Alexa647 or RG7787-Alexa647 and processed as previously described (17 (link)). Fluorescence intensity was analyzed on a FACSCalibur.
Protocol detail
Flow Cytometric Analysis of Antibody Binding
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Corresponding Organization :
Other organizations : MRC Laboratory of Molecular Biology, KU Leuven, Roche (Switzerland)
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Variable analysis
independent variables
- Incubation time (30, 60, and 120 min)
- Fluorescence intensity
- Incubation temperature (37°C)
- Concentration of SS1P-Alexa647 or RG7787-Alexa647 (2 μg/ml)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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