Anti-GFP Antibody [1F1] (A85301)

Transfected HEK293 cells which overexpress GFP protein were stained with Anti-GFP Antibody (1:2,000). Cells which are transfected with GFP are bright green in the left panel. Staining with Anti-GFP Antibody is shown in red in the middle panel, and this is overlaid with the GFP expression in the third panel, so cells appear orange. Most HEK293 cells are not transfected so only the nucleus of these cells can be visualized with a blue DNA stain.

Immunofluorescent analysis of HEK293 cells transfected with a GFP construct, in green, and stained with Anti-GFP Antibody [1F1] (A85301), at a dilution of 1:1,000, in red. The nuclear DNA is visualised in blue using Hoechst staining. The Anti-GFP Antibody [1F1] (A85301) binds to GFP protein expressed only in transfected cells, and as a result cells appear an orange-golden colour.

Western blot analysis of transfected and control HEK293 cell lysates using Anti-GFP Antibody [1F1] (A85301), in green, at a dilution of 1:1,000. The lanes contain samples of: [1] Protein standards, [2] Control, non-transfected cells, [3] cells transfected with an mCherry red fluorescent protein construct and [4] cells transfected with GFP construct. The strong green band at approximately 27 kDa corresponds to GFP protein detected only in cells transfected with GFP construct. The antibody does not bind to mCherry. The same blot was simultaneously probed with Anti-Heat Shock Protein 60 Antibody (A85438), at a dilution of 1:10,000, in red. The single band at 60 kDa represents the HSP60 protein expressed in all preparations, which acts as a positive loading control.

Blot of crude homogenate from HEK293 cells transfected with pFin-EF1-GFP vector (Lane 1) and non-transfected HEK293 (Lane 2) was probed with Anti-GFP Antibody. The pFin-EF1-GFP vector expresses full length GFP. There is a strong clean band at about 27kDa in transfected cells corresponding to GFP which is absent from non-transfected cells.