Invitrogen™ GFP Polyclonal Antibody, Alexa Fluor™ 555

Rabbit Polyclonal Antibody

At the time of preparation, the products are certified to be free of unconjugated dyes and are tested in a cytological experiment to ensure low nonspecific staining. Several Alexa Fluor™ dye-conjugates made from the rabbit anti-GFP IgG fraction are also available. The Alexa Fluor™ dyes provide for extraordinarily bright antibody conjugates. The approximate fluorescence excitation (Ex) and emission maxima for this Alexa Fluor™ 555 conjugate are 555 nm and 565 nm, respectively. These anti-GFP antibody conjugates are suited for detection of native GFP, GFP variants, and most GFP-fusion proteins by western blot analysis and immunocytochemistry. For initial experiments, we recommend trying dilutions ranging from 1:200 to 1:2,000 with our fluorophore-labeled antibodies, a final concentration of 1-10 μg/mL should be satisfactory for most immunocytochemical applications. If using the rabbit anti-GFP antibody-Alexa Fluor™ conjugates in western blotting, dilute the conjugate 1:2,000 in PBS to obtain a final antibody concentration of 1 μg/mL. HeLa and U2-OS cells were tested with this antibody in immunocytochemistry, but not with paraffin-embedded sections. It is a good practice to centrifuge the protein conjugate solutions briefly in a microcentrifuge before use; add only the supernatant to the experiment. This step eliminates any protein aggregates that may have formed during storage, thereby reducing nonspecific background staining.