Does GFP need excitation?
GFP can be excited by the 488 nm laser line and is optimally detected at 510 nm.
How does GFP emit green light?
Solutions of purified GFP look yellow under typical room lights, but when taken outdoors in sunlight, they glow with a bright green color. The protein absorbs ultraviolet light from the sunlight, and then emits it as lower-energy green light.
What are the excitation and emission peaks of GFP?
The GFP from A. victoria has a major excitation peak at a wavelength of 395 nm and a minor one at 475 nm. Its emission peak is at 509 nm, which is in the lower green portion of the visible spectrum. The fluorescence quantum yield (QY) of GFP is 0.79.
How does GFP tagging work?
GFP-tagging is a way of preparing a sample for fluorescence microscopy by using the GFP as a fluorescent protein reporter. This is done by cloning the GFP in frame with the target protein at either the N- or C-terminus of the amino acid chain.
What is GFP excitation?
The GFP from A. victoria has a major excitation peak at a wavelength of 395 nm and a minor one at 475 nm. Its emission peak is at 509 nm, which is in the lower green portion of the visible spectrum.
What does excitation wavelength mean?
Excitation spectra. A fluorophore is excited most efficiently by light of a particular wavelength. This wavelength is the excitation maximum for the fluorophore. This wavelength is the emission maximum for that fluorophore. The excited fluorophore can also emit light at wavelengths near the emission maximum, as shown.
How do you choose excitation and emission wavelength?
In order to achieve maximum fluorescence intensity, the fluorochrome is usually excited at the wavelength at the peak of the excitation curve, and the emission detection is selected at the peak wavelength (or other wavelengths chosen by the observer) of the emission curve.
What is mEGFP?
mEGFP is a basic (constitutively fluorescent) green fluorescent protein published in 2002, derived from Aequorea victoria. It has moderate acid sensitivity.