Confocal microscopy images of COS - 7 cells transfected with expression constructs encoding membrane - tethered EGFP (membrane - EGFP ; top) or nuclear Polycomb 2 - EYFP fusion protein (Pc2 - EYFP ; bottom). The natural fluorescence of the produced proteins is shown in the green channel (left), GFP polyclonal antibody (Cat # PAB1264) signal was detected in the red channel (right). The system was carefully tested for overlap of these two optical channels and images were scanned separately in sequential scanning mode. The blue nuclear stain is also shown.
Immunoprecipitation of GFP-NLS from HEK293 cells using GFP polyclonal antibody (Cat # PAB1264). HEK293 cells were transfected with expression construct encoding GFP-NLS protein. Twenty hours post transfection cells were lysed in non-denaturating conditions (Lysis buffer: 20 mM Tris, pH 7.5, 100 mM NaCl, 0.5% Triton X-100, inhibitors of proteases). Aliquots of cell lysate were immunoprecipitated using a polyclonal GFP polyclonal antibody (Lane 2) or a pre-immune rabbit serum (Lane 3). Immunoprecipitates together with a sample of the cell lysate (Lane 1) were separated on SDS-PAGE polyacrylamide gel and immunoblotted with the GFP polyclonal antibody. The positions of molecular weight markers in kDa are indicated at the left.