Highlights
G1S Cell Cycle Phase Marker Assay for dynamic, non-toxic reporting of cell cycle status in living cells.
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Ad-A-Gene Vectors
This system comprises a range of ready to use recombinant adenoviral preparations. Each adenovirus contains the gene encoding a protein target fused to EGFP or emerald FP, or a response element controlling the expression of the reporter gene, nitroreductase (NTR).
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Green fluorescent Proteins (GFPs)
Bringing Drug Discovery To Life
GE Healthcare has a broad product offering based on Aequorea victoria GFP consisting of both licenses and reagents:
GFP Licenses: a broad portfolio of licenses to the complete range of "red-shifted" and "folding" mutations from Amersham Biosciences, Invitrogen IP Holdings Inc. and BioImage A/S, available from a single source.
Columbia GFP License: a license to the intellectual property owned by Columbia University covering the use of Aequorea victoria GFP.
GFP Assays: cell lines stably transfected with GFP constructs allowing study of translocation events in live-cell assays.
GFP is used in a very broad range of application areas and impacts on all parts of cell biology from the single cell to the whole animal
- Functional genomics:
Use GFPs in: Protein translocation studies, Gene expression studies, Effect of known or unknown genes on GFP fusion proteins, Link to unknown cDNAs & monitor location, translocation & interactions of protein products
Why Choose AvGFP?
- Well established, proven characteristics, extensively documented and published
- Color variants available - Blue, Green, Yellow and Cyan
- Monomeric – non aggregating, ensures accurate reporting of biological activity
- Performance enhancing mutations introduced
- Red shifted mutation S65T for excitation at 488nm, optimal for standard fluorescence instruments
- Fast folding mutation F64L for brighter fluorescence at biological temperatures
- Suitable for use with almost any fluorescence microscope, fluorescence plate reader or fluorescence imager.
- Wide range of validated reagents and assays available
Background
Green fluorescent proteins (GFP) are intrinsically fluorescent proteins derived from bioluminescent marine organisms. GFP comprise a family of evolutionarily related proteins found in animals such as jellyfish and corals that belong to the phylum Cnidaria (stinging aquatic invertebrates). These organisms have evolved GFP as a part of a mechanism that enables them to glow in dark ocean waters. In recent years, scientists have begun to exploit GFP as a tool to explore the internal workings of living cells. The advent of GFP technology has revolutionized cell biology by enabling researchers to visualize proteins within living cells without the need for chemical staining.
First isolated from jellyfish in 1994, Aequorea Victoria Green Fluorecent Protein (AvGFP) is one of a family of fluorescent proteins that have become widely used tools in many biological research applications. AvGFP is a 238 amino acid, 27,000 Dalton monomer, it is intrinsically fluorescent and needs no additional substrates or co factors for fluorescent expression of the protein. This is a big advantage over fluorescent dyes where the protein must be labeled and then delivered to the cell in a fluorescent form, or luciferases which require substrates. When introduced into cellular DNA and expressed as a fusion product with a specific protein, the location and translation of that protein may be tracked, due to the intrinsic fluorescence of the fluorescent protein rights.
AvGFPs can be used in conjunction with almost any fluorescence microscope, fluorescence plate reader, FACS or fluorescence imager. The application for AvGFP, and the information that can be obtained is dependent on the type of detection instrument.
At its most powerful, AvGFP has the capacity to report distribution and redistribution of cellular proteins in automated drug discovery and development applications but this has been hampered to date by the lack of high throughput microscopy platforms allowing sub-cellular resolution. The IN Cell Analyzer 3000 and IN Cell Analyzer 1000 with their ability to image at sub-cellular resolution and at high throughput will be able to exploit the GFP technology to its full potential. Standard fluorimeters and plate readers allow screeners to determine whether a drug candidate causes an increase or decrease in the level of the target. However, it may not be the physical level of the target that is important – rather it may be the localization of the target within the cell in order for it to be interact with further proteins in the cascade.
Various performance enhancing mutations have been introduced such that AvGFP has become the benchmark tool for monitoring live cellular events and interactions in a non destructive way. Use of these commercially available variants of AvGFP is governed by an array of intellectual property. After a series of strategic alliances, GE Healthcare is able to offer access to many of these patents from a single source for a wide range of applications. For more information, e-mail GE Healthcare at gfp@amersham.com, or contact your local area representative.
BioImage is a Danish biotech company specializing in developing drug candidates that exert their activity through modulation of protein translocation. For more information visit their website at www.bioimage.dk. |
