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.
Epitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance or poorly immunogenic proteins when protein specific antibodies are not available. Green Fluorescent Protein (GFP), from the jellyfish Aequorea victoria, has quickly become a versatile marker for monitoring physiological processes, visualizing protein localization, and detecting transgenic expression, and a powerful research tool for assessing gene expression and subcellular protein distribution in fixed or living cells. GFP is excited by and brightly fluoresces when exposed to UV or blue light. This feature makes it ideal as a marker for use in fluorescence microscopy, cytometry, tagging fusion proteins, and assaying transcriptional regulation from gene promoters in vivo. Numerous GFP variants with enhanced and shifted emission spectra (blue, green, and yellow) have been developed through amino acid substitutions at specific residues.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: eGFP; GFP; GFP tag; GFP2; Green Fluoresecent Protein; Turbo GFP