Application
Flow Cytometry
CHO cell line expressing membrane bound human TNFα (stable transfectants) was incubated with 10μg/ml Remicade (anti-human TNFa monoclonal antibody) for 1 h in 4°C. The unbound antibody was washed off by centrifugation (300x g for 5min) and binding of remicade was detected with PE Goat F(ab)2 anti-hIgG Fc (ab98596) – 1:100 (5ug/ml), 30 min incubation in 4°C. The cells were washed twice in FACS buffer (2.5% BSA, 0.1% sodium azide in dPBS), before flow cytometric analysis.
PE goat F(ab)2 anti-hIgG detected binding of remicade to TNFα CHO cell line giving strong positive signal, however there was some non-specific binding to the cells alone. Further optimisation of the reagent concentration and washing procedure should improve the background signal.
PE goat F(ab)2 anti-hIgG detected binding of remicade to TNFα CHO cell line giving strong positive signal, however there was some non-specific binding to the cells alone. Further optimisation of the reagent concentration and washing procedure should improve the background signal.
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提交于 Apr 25 2017