What is the emission wavelength of FITC?
approximately 495 nm and 519 nm
FITC has excitation and emission spectrum peak wavelengths of approximately 495 nm and 519 nm, giving it a green color.
What is fluorescence excitation maximum?
The excitation maximum of the fluorophore is achieved (1) as the energy level of the molecule peaks during the excitation process. As light is emitted from the fluorophore during the fluorescence process energy is lost (2) which results in shift in the emission maximum (3). This process is called the Stokes Shift.
What is emission maximum?
The emission maximum is chosen and only emission light at that wavelength is allowed to pass to the detector. Excitation is induced (usually by means of a monochromator) at various excitation wavelengths and the intensity of the emitted fluorescence is measured as a function of wavelength.
Is fluorescein same as FITC?
Fluorescein derivatives are the most common fluorescent reagents for biological research because of their high absorptivity, excellent fluorescence quantum yield, and good water solubility.
At what wavelength does GFP fluorescence?
Green fluorescent protein (GFP) is the most commonly used fluorescent reporter. Initially isolated from the jellyfish Aqueoria victoria, GFP has since been engineered to generate the brighter and more photostable enhanced GFP (eGFP), with an excitation wavelength of 488 nm and emission wavelength of 509 nm.
How stable is FITC?
KPL’s FITC conjugates are equally stable when stored at either 4°C in lyophilized form or stored frozen after rehydration with reagent quality water. A high level of performance is seen with FITC conjugates that have been stored for up to six years.
What is fluorescence excitation and emission?
A fluorescence emission spectrum is when the excitation wavelength is fixed and the emission wavelength is scanned to get a plot of intensity vs. emission wavelength. A fluorescence excitation spectrum is when the emission wavelength is fixed and the excitation monochromator wavelength is scanned.
What is the difference between fluorescence and emission?
Emission is the process that creates a photon and takes the the atom or molecule in an excited state back to the ground state. The Emission Spectra of H, He and Hg. Fluorescence is the process that first consumes a photon and puts the atom or molecule in an excited state…
Are FITC and GFP the same?
Available Options. Green Fluorescent Protein (GFP) Antibody (FITC) is a Rabbit Polyclonal antibody conjugated to FITC against GFP.
What wavelength does GFP absorb?
395 nm
A mechanism for the fluorophore formation has been proposed (3) but needs to be confirmed by further studies. GFP absorbs blue light at 395 nm, with a smaller peak at 475 nm, and emits green light at 508 nm with a quantum yield of 0.72–0.85 (12, 13).
What is the excitation and emission spectrum of FITC I?
Excitation (orange) and emission (blue) spectrum of FITC isomer I. The excitation maximum is 495 nm and the emission maximum is approximately 519 nm. Depending on the isomer, FITC contains an isothiocyanate group at either position 5 (isomer I) or position 6 (isomer 2) of the bottom benzene ring, see figure 2.
What is the maximum absorbance of FITC?
In the case of FITC, its maximum absorbance falls within the blue spectrum. Therefore, the blue 488 nm laser, which is close to FITC’s absorbance peak of 490 nm, is commonly used to excite this fluorophore. FITC emits fluorescence from 475 to 650 nm, peaking at 525 nm, which falls in the green spectrum.
What does the FITC spectrum look like?
However, FITC’s emission is not restricted to green photons; it also emits yellow, orange, red, albeit at lower probabilities. You may notice something curious about the spectrum if you look closely at the emission curve immediately around the excitation line.
Is the PE signal in the FITC PMT independent of laser line?
Any PE signal you see in the FITC PMT is independent of the laser line. In other words, you would see this signal regardless of whether you excited PE with 488 laser light (the emission in this case would be higher in wavelength than the excitation line) or the 561 nm laser line (the emission would be lower in wavelength than the excitation line).