Rabbit anti-FLAG Tag Recombinant Monoclonal Antibody(S-589-8)

ICC shows positive staining in Histone H3-Flag-V5 tag transfected HeLa cells (top panel) and negative staining in vector-transfected HeLa cells (below panel). Anti-FLAG Tag antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).

WB result of FLAG Tag Rabbit mAb Primary antibody: FLAG Tag Rabbit mAb at 1/1000 dilution Lane 1: 293-F whole cell lysate 20 ug Lane 2: 293-F transfected with Flag tagged GFP whole cell Lysate 20 ug Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 27 kDa Observed MW: 32 kDa Exposure time: 90 s

WB result of FLAG Tag Rabbit mAb Primary antibody: FLAG Tag Rabbit mAb at 1/1000 dilution Lane 1: Nicotiana benthamiana leaves (transfected with HA tagged GFP) lysate 20 ug Lane 2: Nicotiana benthamiana leaves (transfected with Flag tagged GFP) lysate 20 ug Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 27 kDa Observed MW: 35 kDa Exposure time: 120 s (This blot was developed with high sensitivity substrate)