045846
Green-Red Fluorescent Protein EosFP (Not for Export EU)
Size
£544.00
- SKU:
- 045846
- Purity:
- Serum
- Storage Conditions:
- -20°C
- Supplier:
- United States Biological
- Host:
- Rabbit
- Immunogen:
- Full length recombinant EosFP protein expressed in E. coli
- Format:
- Supplied as a liquid.
- Extra Details:
- Fluorescent proteins have become widely used in a variety of experimental paradigms since the characterization of the first one, Green Fluorescent Protein (GFP) about 20 years ago. Some of these fluorescent proteins have the useful property of changing their emission spectrum from green to red following irradiation with blue or UV light, which allows simple but powerful pulse chase type experiments to be performed using more or less standard fluorescence microscopes. We have expressed in E. coli an purified a protein originally isolated from Lobophyllia hemprichii, a type of stony coral. The protein was described in Wiedenannn et al. (2004) and named EosFP, after Eos, the goddess of dawn in Greek mythology. Upon appropriate irradiation the protein changes emission from 516 nm to 581 nm, which happens to fit very conveniently to typical green (~498 nm) and red (~594 nm) filters on fluorescence microscopes. The emission shift is due to an irreversible covalent modification in the fluorochrome and is dependent on the His residue in the His-Tyr-Gly sequence that produces the fluorescence. The original coral protein was mutagenized to prevent tetramerization and dimerization, a requirement if the protein is to be used for fusion protein generation useful for FRET and similar techniques. A recent advance was the development of CaMPARI, an acronym for “Calcium-Modulated Photoactivatable Ratiometric Integrator” (2). This construct contains the GCaMP Calcium indicator fused to two EosFP domains, and will only transit from green to red when there is a coincidence between Calcium level elevation and the appropriate wavelength of light. This allows functional mapping of cellular activation in real time in appropriate transgenic animals (2). The sequence expressed is identical to that listed in Uniprot entry Q5S6Z9. It can be used to verify the expression of an EosFP containing protein construct or to amplify the EosFP signal in cells or tissues. Applications: Suitable for use in Western Blot, Immunocytochemistry and Immunofluorescence. Other applications have not been tested. Recommended Dilutions: Western Blot: 1:1000-1:5000 Immunofluorescence/Immunocytochemistry: 1:5000 Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
- Shipping Conditions:
- Blue Ice