Immunofluorescent analysis of GFP Tag was performed using H3-GFP construct transfected in HEK-293E cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with GFP Monoclonal Antibody (3E6) (Product # A-11120) at 1:50 dilution and Histone H3 Rabbit Polyclonal Antibody (Product # 711055) at 0.5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 555 (Product # A32727) and Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 647 (Product # A32733) respectively at a dilution of 1:2000 for 45 minutes at room temperature. Panel a (Nuclei: Green) represents GFP Monoclonal Antibody (3E6), a green pseudo-colour has been given after image capture. Panel b (Nuclei: Red) represents Histone H3. Panel c (Nuclei: Blue) represents ProLongTM Diamond Antifade Mountant with DAPI (Product # P36962). Panel d represents the merged image showing the co-localization of nuclear signals in transfected cells. Panel e represents untransfected HEK cells. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supplier Page from Thermo Fisher Scientific for GFP Antibody