All lanes : Anti-SH2D1A/SAP antibody [EPR3168] (ab109120) at 1/1000 dilutionLane 1 : MOLT-4 cell line lysateLane 2 : Jurkat cell line lysateLane 3 : K562 cell line lysateLysates/proteins at 10 µg per lane.
Flow cytomertic analysis of permeablilized Molt-4 cells using ab109120 (red), rabbit IgG (negative, green).
Overlay histogram showing Jurkat cells stained with ab109120 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109120, 1/50) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.