FITC Anti-TNF alpha antibody [EPR21753-109]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - FITC Anti-TNF alpha antibody [EPR21753-109] (AB323680)

Flow cytometry overlay histogram showing RAW264.7 treated with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h (green line) or untreated RAW264.7 stained with ab323680 (magenta line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were incubated in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab323680) (1x 10⁶ in 100μl at 1 μg/ml (1/500)) for 30min at 22°C.

Isotype control antibody Rabbit IgG [EPR25A] FITC was used at the same concentration and conditions as the primary antibody (RAW264.7 treated - black line, RAW264.7 untreated - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.