GFP Polyclonal Antibody, HRP, Invitrogen™

Description

• We offer the anti-GFP antibody HRP as purified rabbit IgG fraction and conjugated to horseradish peroxidase (HRP)
• 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
• This anti-GFP antibody HRP conjugate is 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 for immunocytochemical applications and western blot analysis
• 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
• To prepare 0.2 mg/mL stock solutions, reconstitute the lyophilized antibody in 1 mL of phosphate-buffered saline (PBS), pH 7.4
• Store solution at 2-8°C with the addition of thimerosal to a final concentration of 0.02%
• For prolonged storage after reconstitution, add glycerol to a final concentration of 50% (v/v), aliquot, and store at -20°C
• When stored properly, the solution is stable for approximately three months
• 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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