GFP Polyclonal Antibody, Alexa Fluor™ 555, Invitrogen™

Description

• 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
• The jellyfish Aequorea victoria contains green fluorescent protein (GFP Tag) that emits light in the bioluminescence reaction of the animal
• GFP has been used widely as a reporter protein for gene expression in eukaryotic and prokaryotic organisms, and as a protein tag in cell culture and in multicellular organisms
• As a fusion tag, GFP can be used to localize proteins, to study their movement or to research the dynamics of the subcellular compartments where these proteins are targeted
• GFP technology has revealed considerable new insights in the physiological activities of living cells
• GFP is a 27 kDa monomeric protein, which autocatalytically forms a fluorescent pigment
• The wild type protein absorbs blue light (maximally at 395nm) and emits green light (peak emission 508nm) in the absence of additional proteins, substrates, or co-factors
• GFP fluorescence is stable, species independent and is suitable for a variety of applications
• GFP has been used extensively as a fluorescent tag to monitor gene expressin and protein localization
• Moreover, other applications for GFP include its use in assessing protein protein interactions in the yeast two hybrid system, and in measuring distances between proteins in fluorescence energy transfer (FRET) experiments
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